Institute of Biology, College of Science

University of the Philippines Diliman

Extension Services

SERVICE BROCHURE
Version 2.1
 MCCL Extension Service Website

https://sites.google.com/up.edu.ph/mccl-upd-
extension-services/

Version 2.1 January 2024

Note: This work is not for citation.

Editor: Ranelle Janine L. Asi

Mammalian Cell Culture Laboratory, Rm 268 Institute of Biology,

Ma. Regidor St., University of the Philippines Diliman, Quezon City
Philippines 1101

Previous versions:
2.0 September 2023
1.0 April 2023
 TABLE OF CONTENTS
Foreword  i
The Mammalian Cell Culture Laboratory  ii
A. List of Laboratory Services  1
B. Sample Preparation  3
B.1. What kinds of samples are accepted for testing?  3
B.2. How should clients prepare their samples?  5
B.3. Drying samples before testing  7
B.3.1. MCCL Sample Drying Services  8
B.4. Sample packaging  10
C. Phytochemical Assays  12
C.1. DPPH Free Radical Scavenging Assay  12
C.2. Total Phenolics Content Analysis  14
C.3. Total Flavonoid Content Analysis  16
D. Cell-based Assays  18
D.1. MTT Viability/Cytotoxicity Assay  18
D.2. MTT Photo-documentation  21
D.3. Wound Healing/Scratch Assay  22
D.4. Annexin V-PI Apoptosis/Necrosis Assay  25
D.5. Immunofluorescent Staining Assay  28
D.5.1. Ki-67 staining  29
D.5.2. p53 staining  30
D.5.3. Custom marker staining  30
E. Service Procedures  32
E.1. How can clients request for service?  32
E.2. How long does each procedure take?  33
F. Service Deliverables: Final Reports  36
G. Frequently Asked Questions  40
References  42
Acknowledgements  43
 LIST OF FIGURES
Figure 1. Samples that may be accepted for testing  3
Figure 2. Samples that may NOT be accepted for testing  4
Figure 3. Sample preparation pipeline  6
Figure 4. Sample drying and reconstitution  7
Figure 5. Recommendations for sample packaging  10
Figure 6. Sample label format  11
Figure 7. Principle behind the DPPH free radical scavenging assay  12
Figure 8. Principle behind the Folin-Ciocalteu method for
measuring total phenolic content  14
Figure 9. Principle behind the aluminum chloride method for
measuring total flavonoids content  16
Figure 10. Principle behind the MTT viability/cytotoxicity assay  18
Figure 11. Example of phase-contrast images for MTT assay
photo-documentation  21
Figure 12. Principle behind the wound healing/scratch assay  22
Figure 13. Principle behind the Annexin V-PI apoptosis/
necrosis assay  25
Figure 14. Principle behind the indirect method for
immunofluorescent staining assay  28
 LIST OF TABLES
Table 1. Summary of the three sample drying methods  9
Table 2. Service Request Classification for each Sample Type  32
Table 3. Timetable for Sample Drying Procedures  34
Table 4. Timetable for Regular Assay and Confirmatory
Bioassay Procedures  35
Table 5. Summary of MCCL Service Deliverables and
Final Report Contents  36
 Foreword

Ahmad Reza F. Mazahery, Ph.D.

Laboratory Head and Principal Investigator
Mammalian Cell Culture Laboratory
Asst. Prof., Institute of Biology UP Diliman

The Mammalian Cell Culture Laboratory (MCCL) of the Institute of Biology started as a
research facility for testing natural products that had potential therapeutic effects against cancer.
Over the past years under the leadership of now-retired Dr. Sonia D. Jacinto, MCCL has built on
the facilities and capabilities to chemically purify natural product extracts and test them on cell
culture models of cancer. Today, beyond cancer cells, MCCL conducts research in many other
diseases, namely HIV-related chronic complications, tuberculosis, acute myocardial infarction,
and leptospirosis using advanced disease models such as induced pluripotent stem cells.
All of these are made possible through the help of our partners and funding agencies. The
continued support from our funding agencies such as the Department of Science and Technology
(DOST), and the Commission on Higher Education (CHED) allowed us to afford many of the
equipment, personnel, and other logistical support for us to implement these research activities.
We have also established local and international research partnerships that allow us to explore
more relevant diseases.
With its growing research capacities, MCCL also aims to extend its successes to the Filipino
people by catering to the needs of local health researchers outside our institute. Through the
years, MCCL has provided extension services that allowed students and other researchers to
collect scientific data by submitting their samples to our laboratory for testing. With the new
enhancements and continued government support, MCCL has improved its services from the
standard cytotoxicity assays and antioxidant testing as we now offer more stringent biomolecular
analysis and even bioimaging. More than that, we are now also providing scheduled mentoring
activities aimed to provide the needed skills for Filipino health researchers to perform natural
product extraction, basic cell culture techniques, and other relevant biomolecular assays.
Through this, we hope to foster and promote further health research across the country. We also
wish to contribute to the need of a stronger health research labor force.
To help our potential clients and mentees who are interested in availing our services, we have
authored this Service Brochure. In it, you will find detailed explanation of every laboratory service
our laboratory is offering. It will also serve as a guide on how the client can choose the
appropriate service assay for their specific research and what to expect from our services.
Moreover, this brochure also contains the detailed instructions for all needed administrative and
documentary requirements of each service. We hope that all these information will be helpful for
those who wish to avail our extension services.

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The Mammalian Cell Culture Laboratory

The Mammalian Cell Culture Laboratory (MCCL) was

founded in 2011 by then Institute of Biology Professor, Dr.
Sonia D. Jacinto (right photo), who has peerless passion for
studying the potential of Philippine plant natural products
in targeting cancers, inflammatory diseases and other
globally important diseases. The laboratory, which was
initially an academic research site, grew throughout the
years with the exceptional efforts of Dr. Jacinto, her
research team and various research students to eventually
become acknowledged by the Department of Science and
Technology (DOST) as a National Bioassay Facility.
Equipped with the right physical, technical and personnel
capacity, MCCL began to extend services to the Filipino
public by offering bioassay services which can benefit other
health researchers, biologists, chemists and, most
importantly, young research students with no access to cell
culture facilities.
Today, MCCL continues to ardently push its efforts in
scientific research and public service with the leadership of
Dr. Jacinto’s former student and now Institute of Biology
Assistant Professor Dr. Ahmad Reza F. Mazahery and his
research team, with the significant support from the UP
Institute of Biology, DOST and the Commission on Higher
Education (CHED). As of 2023, MCCL continues to increase
its capacity for health research and bioassay services in
terms of natural product testing, cell culture and molecular
analysis, with the commitment of bettering the lives of the
Filipino people.

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A. List of Laboratory Services

 Sample Drying Services

This service can be availed for wet samples, e.g. plant extracts dissolved in
extraction solvents such as ethanol, water, hexane, etc. Sample drying may be
performed through either freeze-drying (lyophilization), rotary evaporation
(rotavap), oven-drying at 40°C, or the combination of any 2 or all 3 methods
(depending on the type, volume and characteristics of the sample). For the
preparation of wet samples to be subjected to the bioassays listed below, this step is
required.

 DPPH Free Radical Scavenging Assay

A phytochemical assay which measures free radical scavenging capacity through
reduction of the DPPH molecule. This service can be availed to assess whether your
submitted sample has antioxidant properties. This service includes testing of 8 fixed
concentrations of your sample, which are to be used to compute for an IC50 value.

 Total Phenolics Content (TPC) Assay

A phytochemical assay which uses the Folin-Ciocalteu method to measure the
concentration of total phenolic compounds in your submitted sample using gallic
acid as standard.

 Total Flavonoid Content (TFC) Assay

A phytochemical assay which uses the aluminum chloride method to measure the
concentration of total flavonoid compounds in your submitted sample using
quercetin as standard.

 MTT Cytotoxicity Assay

A cell-based assay that is used to measure in vitro cellular metabolic activity though
the reduction of the MTT molecule. This service can be availed to assess whether
your submitted sample can induce changes in cell viability through proliferation or
cytotoxic effects after 72 hours of incubation. This service includes testing of 8 fixed
concentrations of your sample, which are to be used to compute for an IC50 value.

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 List of Laboratory Services

 MTT Assay Photo-documentation

This is an additional service for the MTT assay wherein the cells treated with your
submitted sample will be photographed under a phase-contrast microscope. This
service can be availed to assess whether your submitted sample can induce changes
in cell morphology.

 Wound Healing/Scratch Assay

A cell-based assay that is used to measure collective cell migration by documenting
the closure of a wound/scratch made on a cell monolayer over a period of 72 hours.
This service can be availed to assess whether your submitted sample can induce
changes in the rate of wound closure/cell migration. We require availing the MTT
cytotoxicity assay prior to this service in order to establish the best concentration to
test.

 Annexin V-PI Apoptosis Assay

A cell-based assay that is used as a confirmatory step for an observed cytotoxic
effect. This assay detects whether dying cells are undergoing apoptosis or necrosis
by fluorescently tagging cells with Annexin V (green) and propidium iodide (PI) (red).
We require availing the MTT cytotoxicity assay prior to this service in order to
establish the best concentration to test.

 Immunofluorescent Staining Assay

A cell-based assay that is used as a confirmatory step for an observed cellular effect.
This assay detects the presence/absence and cellular localization of a particular
antigen/protein of interest through the use of a fluorescently-conjugated antibody
that specifically binds to that antigen/protein. For our service, we offer
immunofluorescent staining of two well-studied proteins: Ki-67 (a cancer
proliferation marker) and p53 (a tumor suppressor marker). We require availing the
MTT cytotoxicity assay prior to this service in order to establish the best
concentration to test.

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B. Sample Preparation

What kinds of samples

B.1.
are accepted for testing?
We offer testing of extracts/compounds from any natural
product source, and semi-pure/pure compounds.
WE ACCEPT samples that have been processed through crude extraction (such as
aqueous extraction, liquid-liquid solvent extraction, distillation, etc.), samples that
are semi-purified (such as chromatographic fractions), and samples that are pure
compounds (such as drug compounds and novel compound isolates).

WET
SAMPLES
Drugs / pure
compounds

DRIED SAMPLES

*Crude extracts from plants/ Chromatographic **Lyophilized powders /

microorganisms/animal sources fractions compounds

*Wet samples such as crude extracts

still dissolved in their extraction **Lyophilized
solvents can be submitted for drying unprocessed
services prior to bioassay testing. plant parts,
herbs, fruits,
etc. will NOT
be accepted

Figure 1. Samples that may be accepted for testing.

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 Sample Preparation

B.1.What kinds of samples

are accepted for testing?
There are limitations for the type of samples that can be
feasibly tested through our bioassay services.

WE DO NOT ACCEPT samples that are unprocessed and cannot be processed or

tested through our available services, samples with risk of bio-contamination, and
samples that are not readily dissolvable in our assay reagents.

rock/soil/non-
Food samples dissolvable powders

Wood/bark samples Live micro- Metallic

Whole unprocessed
organisms objects
plants/plant parts

WE DO NOT OFFER ANY FORM

A bioassay plate is OF EXTRACTION SERVICE
comprised of very small for these kinds of samples.
wells which can only fit
dissolvable crude
extracts, semi-purified to
purified compounds, and
drug compounds

Figure 2. Samples that will NOT be accepted for testing.

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Sample Preparation

How should clients

B.2.
prepare their samples?
MCCL does not offer extraction services hence the client is
responsible for performing crude extraction/isolation procedures prior to
submitting their sample. The workflow of sample preparation is
summarized in Figure 1 (next page), specifying which procedures are the
responsibility of either the client (blue) or MCCL (maroon).

The choice of crude extraction/isolation procedure is entirely

dependent on the client. The client should discuss the selection of these
procedures with their research team/adviser/supervisor. MCCL is not
responsible for providing technical advice or recommending any
specific extraction procedure. For more information on how to perform
natural product extraction, you may review the comprehensive review
article by Zhang et al. (2018).

How much of each sample should be submitted?

MCCL will only accept a certain range of volume or weight of sample

depending on the Service Requested. Please refer to Table 2 on page 32.

We recommend sending the sample/s at a weight/volume higher than

the minimum limits to ensure appreciable sample amounts for retests.
However, samples should not exceed the maximum limits to ensure faster
processing.

5
 Sample Preparation

How should clients

B.2.
prepare their samples?
CLIENT CLIENT CLIENT

WHOLE CRUDE EXTRACTION ISOLATION / PURIFICATION

SPECIMEN
Wet Crude Extract Wet Fractions

Legend:
CLIENT OR MCCL
Sample will be
accepted
Sample will NOT
be accepted
Required step
Optional step

CLIENT SAMPLE DRYING MCCL

Dry Sample

SAMPLE PICK-UP ASSAY SERVICES

Figure 3. Sample preparation pipeline.

6
Sample Preparation

B.3. Drying samples before

testing
Why do we need to dry samples before testing them
in our bioassays?

We need the accurate weight of the extracted compounds.

Common extraction methods use carrier solvents to extract certain groups of
compounds (e.g., alkaloids flavonoids, phenols, etc.). These compounds cannot
be directly quantified while dissolved in solvents. Most of these compounds are
non-volatile and will precipitate in solid forms (powder, colloidal mixtures, etc.)
after drying. These can then be accurately weighed and reconstituted at right
concentrations using new solvents appropriate for the requested bioassay.

We must remove the extraction solvent because it may

interfere with the assay reactions.
Most extraction methods use solvents that are directly cytotoxic to cells and/or
may interfere with chemical reactions during the assay. We must completely
remove these solvents and reconstitute the extracted compounds in the right
assay solvents which also serve as our negative controls.

Figure 4. Sample drying and reconstitution. Sample drying may be

performed either by the client or through MCCL drying service.
Reconstitution in the assay solvent will be performed by MCCL.

7
 Sample Preparation
Sample Preparation

B.3. Drying samples before

testing
SERVICE B.3.1. MCCL Sample Drying Services
Clients can either dry their own samples using their preferred methods
prior to sample submission or avail of the drying services offered by MCCL.
There are three methods available for our sample drying service:

1. Freeze-drying 2. Rotary Evaporation 3. Oven drying

(lyophilization) (Rotavap) at 40°C

Each method has its own advantages and disadvantages and may only
be applicable to certain types of samples. It order to fully dry your wet
sample, it may take the combination of any 2 or all 3 of the methods listed
above. Table 1 (next page) lists the known advantages and disadvantages
for each method.
In order to avail of our drying services, the client must also indicate the
extraction solvent used so we can assess if the drying service method is
appropriate for the wet sample. If the client choses an inappropriate drying
method, MCCL will inform them of any changes to the method prior to the
acceptance of their samples.

8
Sample Preparation

B.3. Drying samples before

testing
Table 1. Summary of the three sample drying methods for Wet Samples.
Standard Protocol Advantages Disadvantages Applicable Samples
1. Sample will be stored • Samples are protec- • Higher costs, longer Samples dissolved
overnight at –80°C ted from high tempe- time, and more steps in high percentage of
ratures that cause for sample water (>95% of total
2. Frozen sample will be volume)
loaded into appropriate degradation of preparation
certain compounds
containers and loaded
like proteins • Cannot be used to dry Samples dissolved
in the freeze-dryer samples containing in organic solvents
chamber • Highest efficiency in solvents with freezing (e.g. ethanol, hexane,
preserving nutrients, points below -55°C ethers, etc.)
3. The machine will be
run at –50-55°C, 0.5-1 proteins and other such as alcohols and Oily/colloidal
mbar until the sample biomolecules organic solvents samples
is visibly dry (but not • Several samples in
more than 6 hrs) one run

1. The cooling system will • Quickest and most • Will subject samples Samples dissolved
be set to 4°C and the efficient drying to moderately high in any percentage of
water bath will be set method for large temperatures which water
to 40°C volume of liquid may cause degrada- Samples dissolved
2. Sample will be loaded samples tion of heat-sensitive in organic solvents
and run in the rotavap compounds like (e.g. ethanol, hexane,
machine until a signi- proteins ethers, etc.)
ficant reduction in • One sample per run Oily/colloidal
solvent is observed
• Samples must further samples
3. Before turning viscous/ be dried in an oven to
thick, sample will be fully remove any
transferred to an oven remaining solvent
set at 40°C

1. Sample will be loaded • Cheapest method for • Will subject samples Samples dissolved
into a glass petridish sample drying to moderately high in any percentage of
temperatures which water
2. Sample will be placed • Can be used for a may cause degrada-
inside the oven-drier much wider variety of Samples dissolved
set at 40°C tion of heat-sensitive in organic solvents
samples compounds like (e.g. ethanol, hexane,
3. Sample will be • Several samples in proteins ethers, etc.)
incubated until all one run
solvents are fully Oily/colloidal
removed samples

9
 Sample Preparation

B.4. Sample packaging

REMINDER: The client must first undergo the steps of applying for Service
Request detailed in Section E prior to submitting samples to MCCL. Walk-
in sample submissions and submissions without labels, due procedure and
confirmed schedules will automatically be rejected.

After performing extraction/isolation procedures, the client is

responsible for appropriately and securely packing their sample before
submitting it to MCCL. The choice of packaging material is entirely
dependent on the client, however the following guidelines may be
considered to ensure safe sample delivery:
RECOMMENDED

Stored in lab-grade Labelled with Caps tightly Covered with Covered with
bottles or plastic tape & water- secured with aluminum foil bubble wrap
containers with resistant ink parafilm wrap to protect from to prevent
tight caps UV/light breakages

NOT RECOMMENDED

Fragile/open- plastic/paper water/soda loosely sealed unclear

mouth glass bags/boxes bottles containers labels
containers

Figure 5. Recommendations for sample packaging

10
Sample Preparation

B.4. Sample packaging

The client MUST securely attach a label to the sample bottle/vial/tube
prior to delivery. The label must contain the following details:
• Client Name (as indicated in Service Form)
• Sample Name (as indicated in Service Form)
• Solvent used (indicate percent if mixed solvent)
• Assays Requested (as indicated in Service Form)
• Storage Temperature (in degrees Celsius)

EXAMPLE:

Figure 6. Sample label format

REMINDER: Samples with improper labels or without labels will

automatically be rejected.

11
C. Phytochemical Assays

SERVICE C.1. DPPH Free Radical

Scavenging Assay
The DPPH Free Radical Scavenging Assay measures antioxidant
activity through a colorimetric chemical reaction wherein the free radical
molecule, DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) is reduced in
the presence of an antioxidant (Figure 7). When DPPH is dissolved in
ethanol, a purple solution is produced. When an antioxidant is added to this
solution, it donates a hydrogen molecule to DPPH to form diphenylpicryl
hydrazine (DPPH-H) which forms a yellow solution in ethanol. This change
in color is then measured using a spectrophotometric reader set at a
wavelength of 570nm. The positive control for this assay is Gallic Acid, a
strong antioxidant derived from plants, and the negative control is the
sample solvent/diluent.

Concentration of antioxidant

Absorbance reading (570nm)

Figure 7. Principle behind the DPPH free radical scavenging assay

12
 Phytochemical Assays

SERVICE C.1. DPPH Free Radical

Scavenging Assay
The DPPH assay tests how an antioxidant inhibits the formation of (or
“scavenge”) DPPH free radicals. This activity will be computed as percent
inhibition based on the formula below:

Our service will test and compute % inhibition of the sample at six (6)
fixed concentrations in three (3) trials:

Sample concentration, ug/ml

500 250 125 62.5 31.25 15.625 7.8125 3.90625

Based on the % inhibition values of this 2-fold titration scheme, we will

calculate the half-maximal inhibitory concentration (IC50) of the sample
in order to assess the strength of its antioxidant activity based on the
criteria set by Phongpaichit et al. (2007):

IC50 (ug/ml) Antioxidant activity

< 10 Very strong
10-49.99… Strong
50-99.99… Moderate
100-249.99… Weak
>250 Inactive

The positive control, gallic acid, will be tested at fixed concentrations to

ensure the validity of the assay. The negative control is the sample solvent/
diluent which will be tested at % volumes parallel with the eight
concentrations of the sample.
13
Phytochemical Assays

SERVICE C.2. Total Phenolics

Content Assay
One of the methods used for Total Phenolics Content (TPC) Assay is
based on the use of the Folin-Ciocalteu (F-C) reagent. The F-C reagent is
a yellow solution of phosphotungstic and phosphomolybdic acid
compounds. In the presence of phenols in an alkaline condition, these
compounds are reduced, producing a dark blue solution (Figure 8). This
change in color is then measured using a spectrophotometric reader set at a
wavelength of 760nm. Based on these absorbances, the concentration of
total phenolic compounds is interpolated from a standard curve established
using several concentrations of Gallic Acid, a known antioxidant phenolic
compound.

Concentration of phenols

Absorbance reading (760nm)

Figure 8. Principle behind the Folin-Ciocalteu method for

measuring total phenolic content

14
 Phytochemical Assays

SERVICE C.2. Total Phenolics

Content Assay
To establish a standard curve for our TPC assay service, Gallic Acid will
be assayed at concentrations of 0, 10, 100, 500 ug/ml and the resulting
absorbances will be plotted on an XY plane. The linear equation will be
derived as:

y (absorbance) = m * x (concentration) + b

sample

Using the Gallic Acid standard curve, the total phenolic content of the
sample will interpolated and computed as gallic acid equivalent (GAE)
mg/g sample using a formula based on the Beer-Lambert Law of
Absorbance:

TPC (GAE mg/g) = C * DF * V/m

where; C = concentration (gallic acid), mg/ml
DF = dilution factor
V = volume of sample tested, ml
m = mass of sample tested, g

For our TPC assay service, samples will be analyzed at concentrations

of 100, 500 and 1000 ug/ml in three (3) trials.

15
Phytochemical Assays

SERVICE C.3. Total Flavonoids

Content Assay
One of the methods used for Total Flavonoids Content (TFC) Assay
is based on the use of aluminum chloride (AlCl3) solution. AlCl3 forms acid
stable complexes with the keto and/or hydroxyl groups of flavonoid
molecules, producing a purple solution (Figure 9). This change in color is
then measured using a spectrophotometric reader set at a wavelength of
570nm. Based on these absorbances, the concentration of total flavonoid
compounds is interpolated from a standard curve established using several
concentrations of Quercetin, a known antioxidant flavonoid compound.

Concentration of flavonoids

Absorbance reading (570nm)

Figure 9. Principle behind the aluminum chloride method

for measuring total flavonoids content

16
 Phytochemical Assays

SERVICE C.3. Total Flavonoids

Content Assay
To establish a standard curve for our TFC assay service, Quercetin will
be assayed at concentrations of 0, 10, 100, and 500 ug/ml and the resulting
absorbances will be plotted on an XY plane. The linear equation will be
derived as:

y (absorbance) = m * x (concentration) + b

sample

Using the Quercetin standard curve, the total phenolic content of the
sample will interpolated and computed as quercetin equivalent (QE) mg/
g sample using a formula based on the Beer-Lambert Law of Absorbance:

TFC (QE mg/g) = C * DF * V/m

where; C = concentration (quercetin), mg/ml
DF = dilution factor
V = volume of sample tested, ml
m = mass of sample tested, g

For our TFC assay service, samples will be analyzed at concentrations

of 100, 500 and 1000 ug/ml in three (3) trials.

17
D. Cell-based Assays

SERVICE MTT Viability/

D.1.
Cytotoxicity Assay
The MTT viability/cytotoxicity assay measures cellular metabolic
activity as an indicator of viability, proliferation or cytotoxicity. This activity
is quantified through a colorimetric chemical reaction wherein the yellow
compound MTT (3-(4,5-dimethylethylthiazol-2-yl)-2,5-diphenyltetra-
zolium bromide) is reduced by NADH in the presence of cellular enzymes,
leading to the formation of a crystal product called formazan (Figure 10).
When formazan is dissolved in dimethyl sulfoxide (DMSO), it then forms a
deep purple solution. This change in color is then measured using a
spectrophotometric reader set at a wavelength of 570nm. The positive
control for this assay is Doxorubicin, a well-known anthracycline drug
which induces cell death, and the negative control is DMSO, which is the
assay solvent used to dissolve the sample/compound being tested.

Cytotoxicity

Cellular viability

Absorbance reading (570nm)

Figure 10. Principle behind the MTT viability/cytotoxicity assay

18
 Cell-based Assays

SERVICE MTT Viability/

D.1.
Cytotoxicity Assay
The MTT assay tests how a compound/extract inhibits or promotes the
viability of cultured cells after a period of incubation. This activity will be
computed as percent inhibition based on the formula below:

Our service will test and compute % inhibition of the sample at eight
(8) fixed concentrations after 72-hr incubation in three (3) trials:

Sample concentration, ug/ml

100 50 25 12.5 6.25 3.125 1.5625 0.78125

Based on the % inhibition values of this 2-fold titration scheme, we will

calculate the half-maximal inhibitory concentration (IC50) of the sample
in order to assess the strength of its cytotoxic activity based on the criteria
set by Jokhadze et al. (2007):

IC50 (ug/ml) Cytotoxic activity

< 10 Highly Active
10-29.99… Active
30-100 Moderately Active
> 100 Weak to Inactive

In the case that the sample promoted cell proliferation (negative %

inhibition values) or the data points are non-sigmoidal, IC50 values will not
be reported. The IC50 value of the positive control, doxorubicin, will be
established to ensure the validity of the assay. The negative control, DMSO,
will be tested at % volumes parallel to the 8 concentrations of the sample.

19
Cell-based Assays

SERVICE MTT Viability/

D.1.
Cytotoxicity Assay
Selection of Cell Lines for MTT Assay

REMINDER: MCCL DOES NOT SELL CELL LINES. Our cell lines are
available because we are conducting ongoing research projects that make
use of these cell lines. We are not permitted by the original
manufacturers (ATCC, Lonza, Sigma) to resell these biomaterials.

The client may choose one or more of the available cell lines at MCCL
for their requested MTT assay. MCCL is not responsible for providing
technical consultation regarding which cell line/s is/are most appropriate
for the client’s research. A list of available cell lines is posted at the MCCL
Extension Service Website or in our Service Forms, which are
periodically updated based on the availability of active cultures. Below are
the types of cell lines typically available at MCCL for MTT assay:

Human carcinoma cell lines Human metastatic cancer cell lines

(e.g. HCT 116, MCF-7, A549, etc.) (e.g. SH-SY5Y, MDA-MB-123, etc.)

Human blood cancer cell lines Normal cell lines

(e.g. THP-1, H9, etc.) (e.g. HDFn, NIH-3T3, AA8, etc.)

20
Cell-based Assays
Cell-based Assays

SERVICE MTT Assay

D.2.
Photo-documentation
Changes in cellular morphology is an important parameter to assess
the effects of an extract/compound being studied. The MTT assay photo-
documentation service is offered as an additional service since it makes
use of a different equipment (inverted phase-contrast microscope). If this
service is availed along with the MTT assay, representative phase-contrast
photomicrographs of the cultured cells will be taken before and after 72-hr
treatment with the samples and controls.

REMINDER: MTT assay photo-documentation service cannot be availed

AFTER the MTT assay has been completed.

Example:

HCT 116 colorectal cancer cells HCT 116 colorectal cancer cells
before treatment after 72-hr treatment w/ Doxorubicin

Figure 11. Example of phase-contrast images for MTT assay photo-

documentation

21
Cell-based Assays

SERVICE D.3. Wound Healing/

Scratch Assay
The Wound healing assay is a test used to asses whether an extract/
compound can promote or inhibit cell migration on a 2D surface. It is
sometimes also called the Scratch assay because this technique is
performed by making a scratch on the surface of a cell monolayer and
observing how fast the gap closes (Figure 12). Phase-contrast microscopy is
used at different time points to document gap closure and take
photomicrographs for succeeding image analysis. This assay is a valuable
tool especially for research topics involving skin regeneration or cancer
metastasis.

Figure 12. Principle behind the wound healing/scratch assay

22
Cell-based Assays
Cell-based Assays

SERVICE D.3. Wound Healing/

Scratch Assay

The wound healing assay tests how a compound/extract inhibits or

promotes the rate of gap closure of cultured cells after a period of
incubation. This activity will be computed as percent scratch area
(relative to Day 0) and fold change (FC) in wound closure (relative to
the negative control) based on the formulas below:

Our service will test and compute % scratch area and FC (wound
closure) after 3-day incubation in three (3) trials. Representative phase-
contrast images at Day 0, Day 1 and Day 3 will be provided.

Our wound healing assay service allows the client to choose at

which concentration the sample will be tested based on the results of
the MTT cytotoxicity assay prior to this service. NOTE: We recommend a
concentration at which the sample did NOT induce cytotoxic effects. Cell
death will lead to the misinterpretation of the results of the wound healing
assay.

There is no positive control for this assay. The negative control,

DMSO, will be tested at % volume parallel with the concentration of the
sample.

23
Cell-based Assays

SERVICE D.3. Wound Healing/

Scratch Assay

Selection of Cell Lines for Wound Healing/Scratch Assay

REMINDER: MCCL DOES NOT SELL CELL LINES. Our cell lines are
available because we are conducting ongoing research projects that make
use of these cell lines. We are not permitted by the original
manufacturers (ATCC, Lonza, Sigma) to resell these biomaterials.

The client may choose one or more of the available cell lines at MCCL
for their requested wound healing assay. MCCL is not responsible for
providing technical consultation regarding which cell line/s is/are most
appropriate for the client’s research. A list of available cell lines is posted at
the MCCL Extension Service Website or in our Service Forms, which are
periodically updated based on the availability of active cultures. Since the
wound healing assay relies on the principles of cell migration on a flat 2D
surface, the assay is limited to adherent cell lines only. Below are the
types of cell lines typically available for wound/healing assay at MCCL:

Human carcinoma cell lines Normal adherent cell lines

(e.g. HCT 116, MCF-7, A549, etc.) (e.g. HDFn)

24
Cell-based Assays
Cell-based Assays

SERVICE D.4. Annexin V-PI Apoptosis/

Necrosis Assay
The Annexin V-PI Apoptosis/Necrosis assay is a confirmatory test
used to asses whether cells treated with an extract/compound are
undergoing known stages of cell death via Apoptosis and/or Necrosis. In
healthy live cells, a cell membrane component called phosphatidylserine
(Ptd-LSer) is only found at the cytosolic side. During Early Apoptosis, the
membrane flips, exposing Ptd-L-Ser to the cell’s outer side. Annexin V
conjugated with a green fluorescent dye (FITC) can then bind to Ptd-L-Ser
from outside of the cell. During Late Apoptosis, the membrane degrades,
allowing the entry of another red fluorescent dye called propidium iodide
(PI). During Necrosis, the cell membrane has disintegrated and PI tags the
remaining nuclear debris. Based on these principles, live healthy cells with
intact cell membranes will not fluoresce; cells undergoing Early Apoptosis
will exhibit green fluorescence only, cells undergoing Late Apoptosis will
exhibit both green and red fluorescence, and cells undergoing Necrosis will
exhibit red fluorescence only (not shown) (Figure 13). A nuclear fluorescent
stain such as DAPI will be used to stain all cells for quantification.

Figure 13. Principle behind the Annexin V-PI apoptosis/necrosis assay

25
Cell-based Assays

SERVICE D.4. Annexin V-PI Apoptosis/

Necrosis Assay
The Annevin V-PI assay tests whether a compound/extract induces cell
death via the apoptosis pathway. Quantification of this activity will be
performed using an inverted fluorescence phase-contrast microscope
through visual counting. NOTE: MCCL does not offer flow cytometry
analysis or fluorometric reading for this assay. Apoptotic activity will be
computed as % Early Apoptotic cells, % Late Apoptotic cells and %
Necrotic cells based on the formulas below:

Our service will test and compute these % values in three (3) trials.

Our Annexin V-PI apoptosis assay service allows the client to choose
at which concentration the sample will be tested and at which time-
point the assay will be performed based on the results of the MTT
cytotoxicity assay prior to this service. NOTE: We recommend a
concentration at which the sample significantly reduced cell viability and
induced morphological changes.

The positive control for this assay is Doxorubicin, a well-known

anthracycline drug which induces cell death via both apoptosis and necrosis.
The negative control, DMSO, will be tested at % volume parallel with the
concentration of the sample.

26
Cell-based Assays
Cell-based Assays

SERVICE D.4. Annexin V-PI Apoptosis/

Necrosis Assay

Selection of Cell Lines for Annexin V-PI Assay

REMINDER: MCCL DOES NOT SELL CELL LINES. Our cell lines are
available because we are conducting ongoing research projects that make
use of these cell lines. We are not permitted by the original
manufacturers (ATCC, Lonza, Sigma) to resell these biomaterials.

The client may only choose among the cell lines which have previously
been used in the MTT Cytotoxicity/Viability Assay for the sample. MCCL is
not responsible for providing technical consultation regarding which
cell line/s is/are most appropriate for the client’s research.

27
Cell-based Assays

SERVICE D.5. Immunofluorescent

Staining Assay
The Immunofluorescent (IF) staining assay is a confirmatory test
used to asses the presence/absence and cellular localization of a target
antigen/protein of interest through the use of an antibody that specifically
binds to that target. For this service, the Indirect Immunofluorescence
method (Figure 14) will be used. In this method, a primary antibody that
was produced from a different host species (e.g. rabbit, goat, mouse, etc.)
will be used to tag antigens/proteins in a human-derived cell line. Since it
came from a different host, it can then be specifically tagged with a
secondary antibody for that species. This secondary antibody is
conjugated to a fluorochrome (i.e. FITC, Cy3, AlexaFluor, etc.) which can
then be visualized under a fluorescence microscope, ultimately indicating
the presence of the antigen/protein of interest in the cells. A nuclear
fluorescent stain such as DAPI is used to stain all cells for quantification.

Figure 14. Principle behind the indirect method for

immunofluorescent staining assay

28
Cell-based Assays
Cell-based Assays

SERVICE D.5. Immunofluorescent

Staining Assay
For our IF staining assay service, we offer staining of two well-studied
proteins: the proliferation marker Ki-67 and the cell cycle suppressor
p53. We also offer IF staining service for custom markers

D.5.1. Ki-67 staining

The Ki-67 protein is a nuclear protein that is highly expressed in

proliferating cells (above photo) (Sun & Kaufman, 2018). Because there is
abnormally high cell proliferation in cancer tumors, Ki-67 has been gaining
interest as a biomarker for several types of cancers (Li et al., 2015). Hence,
the decrease of this marker may serve as a confirmatory test for extracts/
compounds that have potential anti-cancer properties. At the same time,
the increase of this marker may serve as a confirmatory test for a pro-
proliferative action of extracts/compounds on normal cells.

29
Cell-based Assays

SERVICE D.5. Immunofluorescent

Staining Assay
D.5.2. p53 staining

The p53 protein is a transcription factor that is found in the nucleus

when activated (above photo). This widely-studied protein is known to
control cell proliferation and death (Ozaki & Nakagawara, 2011). The p53
protein binds to DNA to inhibit the expression of cell cycle and apoptotic
genes (Chen, 2016). Moderate expression of p53 is found in normal cells,
but the increase of nuclear p53 serves as a biomarker for cell senescence.
The development of many cancers is associated with the suppression or
dysfunction of p53. Hence, the decrease or complete knockout of p53
expression is considered a sign for cancer progression.

D.5.3. Custom marker staining

The client has the option to request for IF staining assay using a
custom marker. The client MUST provide the primary antibody and
indicate the host species it was produced from in the. MCCL will then
provide the secondary antibody appropriate for the client-provided
antibody.

30
Cell-based Assays
Cell-based Assays

SERVICE D.5. Immunofluorescent

Staining Assay
The IF staining assay detects and roughly quantifies the presence of
specific antigens/proteins. Quantification of these targets will be performed
using an inverted fluorescence phase-contrast microscope through visual
counting. NOTE: MCCL does not offer flow cytometry analysis or
fluorometric reading for this assay. The amount of stained cells will be
computed as percent marker-positive cells (ie. Ki-67+ or p53+) using
the formula:

Our service will test and compute these % values in three (3) trials.

Our IF staining assay service allows the client to choose at which

concentration the sample will be tested and at which time-point the
assay will be performed based on the results of the MTT cytotoxicity
assay prior to this service. NOTE: We recommend a concentration at which
the sample significantly induced changes in cell viability.

The positive control for each marker will be a case-to-case basis,

depending on the cellular effect being investigated. The negative control,
DMSO, will be tested at % volume parallel with the concentration of the
sample.

Selection of Cell Lines for IF Staining Assay

The client may only choose among the cell lines which have previously
been used in the MTT Cytotoxicity/Viability Assay for the sample. MCCL is
not responsible for providing technical consultation regarding which
cell line/s is/are most appropriate for the client’s research.

31
E. Service Procedures

E.1. How can clients request

for service?
Depending on the type of sample/s to be submitted, the client must
first determine the Class of Service Request they will apply for. Refer to
Table 2 below:

Table 2. Service Request Classification for each Sample Type.

Sample
Amount Serviceable Service Request Classification
Type
10–50mL (Freeze-drying)
Wet Class I Sample Drying Services Only
50–500mL (Rotavap/Oven drying)
(in solvent at
an unknown 10–50mL (Freeze-drying) Sample Drying Services followed by
concentration) Class II
50–300mL (Rotavap/Oven drying) Regular Assay Services

Dried 5mg–5g

Diluted 1–5mL at a concentration of Class III Regular Assay Services Only

at a known 10mg/mL1 in DMSO/water/
concentration ethanol2)
Sample previously submitted via Class II or III
Class IV Confirmatory Bioassay Services
Service Request

1
Recommended concentration only. If concentration cannot be adjusted (e.g., sample is a
commercially procured reagent/drug), contact us at mccl.upd@up.edu.ph
2
If your diluent is not DMSO/water/pure ethanol, you must submit 1-5mL of the diluent in
addition to your diluted sample. This will serve as negative control for the assays

After determining the Class of Request, the client must fill-up the
appropriate Service Form (MCCL Form 1) provided in the MCCL
Extension Service Website. Further transactions will then proceed via e-
mail based on our Service Timelines.

32
 Service Procedures

E.2. How long does each

procedure take?
REMINDER: MCCL is not a full-time service facility. All of the research and
administrative staff involved in performing services are employed under
government-funded research projects which are their full-time jobs. Our
extension services are performed on a part-time basis and may take
longer compared to full-time commercial laboratory service centers.

Depending on Class of Service Request, each procedure will take

varying lengths of time to complete. Table 3 (next page) details each step
for our Sample Drying Procedures while Table 4 (page 35) details each
step of the Regular Assay and Confirmatory Bioassay Procedures. NOTE:
These timelines do not take into account the length of time required for
other administrative processes including filing a request, delivering/picking-
up the samples, and paying for the service fees.

MCCL operates similar to a Philippine government agency, wherein a

Citizen’s Charter is released to guide the clients on how long every step of
the entire service will take, from the initiating the Service Request up to the
release of the Final Report/s. Our Citizen’s Charter may be obtained by
downloading it from the MCCL Extension Service Website.

How much is each service and how can I pay?

MCCL service fees may change periodically based on the current

market prices of the reagents and consumables to be used in the requested
service procedure. The most updated List of Service Fees can be
downloaded from the MCCL Extension Service Website.

An official Statement of Account (SOA) and instructions for payment

will only be provided after the client has officially filled a Service Request
and has submitted their samples to MCCL.

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Service Procedures

Table 3. Timetable for Sample Drying Procedures based on MCCL standard protocols.
Sample Drying Time Table
Drying method Service Steps Estimated Time
RA assignment and scheduling 5 days
Sample preparation (ULTF) 1 day
Freeze-drying/ 12-16
lyophilization Lyophilization (for 50mL sample) 1-5 days* days
Report writing, review and endorsement 5 days
RA assignment and scheduling 5 days
Rotary Sample preparation and loading in single run of
Rotavap machine (for 500mL sample) 1-7 days** 12-20
evaporation
Sample collection and further drying in oven days
(Rotavap) 1-3 days**
Report writing, review and endorsement 5 days
RA assignment and scheduling 5 days
Sample preparation and oven drying (for 300mL 11-13
Oven drying sample) 1-3 days**
days
Report writing, review and endorsement 5 days

*depends on the viscosity of the sample

**depends on the total number of samples accepted from all clients in a batch

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 Service Procedures

Table 4. Timetable for Regular Assay and Confirmatory Bioassay Procedures based on
MCCL standard protocols.

Assay Time Table

Assay Service Steps Estimated Time
RA assignment and scheduling 5 days
Regular Assays: Sample preparation and assay performance 2 days
Phytochemical 12 days
(DPPH/TPC/TFC) Data gathering and analysis 3 days
Report writing, review and endorsement 2 days
RA assignment and scheduling 5 days
Cell line revival and expansion 7-14 days*
Cell seeding in assay plates 1 day
Regular Assay:
Sample preparation, treatment of plated cells 1 day 24-31
Cell-based
days
(MTT/Photodoc) Treatment incubation and assay termination 3 days
Data gathering and analysis 5 days
Report writing, review and endorsement 2 days
RA assignment and scheduling 5 days
Cell line expansion 7 days
Confirmatory Cell seeding in assay plates 1 day
Bioassays Sample preparation, treatment of plated cells 1 day
(Annexin V-PI/ 31 days
Treatment incubation and assay termination 3 days
Wound healing/
IF staining) Staining procedure ang image capture 2 days
Data gathering and image analysis 10 days
Report writing, review and endorsement 2 days
*depends on the growth rate stability of selected cell line/s

35
F. Service Deliverables & Final Reports

As the final deliverable, MCCL will release a Final Report specific to

each Service Requested. Please refer to Table 5 (below) for the content of
each report.

Table 5. Summary of MCCL Service Deliverables and Final Report Contents (1 of 4)

Service Deliverables and Final Reports
Other
Services Report Form Report Content Deliverables
• Methodology and no. of hours
Sample MCCL Form 4G - performed Dried
Drying Final Report [Drying] Sample/s
• Percent yield after drying
• Methodology performed
• Table 1. Summary of Percent
DPPH MCCL Form 4D - inhibitions and IC50 (n = 3) N/A
Assay Final Report [DPPH]
• Table 2. Raw absorbance readings at
570nm per trial
• Methodology performed
• Table 1. Summary of average
absorbance readings at 760nm and
computed Gallic Acid equivalents
TPC MCCL Form 4E - (GAE) and TFC of the sample at the
indicated concentrations. N/A
Assay Final Report [TPC]
• Table 2. Summary of blank-adjusted
absorbance readings at 760nm.
• Figure 1. Plots for Gallic acid Standard

• Methodology performed
• Table 1. Summary of average
absorbance readings at 450nm and
computed Quercetin equivalents (QE)
TFC MCCL Form 4F - and TFC of the sample at the indicated
concentrations. N/A
Assay Final Report [TFC]
• Table 2. Summary of blank-adjusted
absorbance readings at 450nm.
• Figure 1. Plots for Quercetin Standard
curve per trial

36
 Service Deliverables & Final Reports

Table 5. cont. (2 of 4)
Service Deliverables and Final Reports, cont.
Other
Services Report Form Report Content Deliverables
• Methodology performed
• Table 1. Percent inhibition computed
from the absorbance readings of each
concentration of the positive control,
doxorubicin, against <cell line>
MTT MCCL Form 4A -
N/A
Assay Final Report [MTT] • Table 2. Percent inhibition computed
from the absorbance readings of each
concentration of the sample against
<cell line>
• Table 3. Absorbance readings
(OD570nm) of treated <cell line>

• Figure 1. A: <cell line> in T-25 culture

flask before seeding. B: <cell line> in 96
-well microplates after seeding prior
treatment.
• Figure 2. A: <cell line> treated with
highest concentration of DMSO (1% v/
v) after 72-hour incubation. B: <cell
MTT line> treated with lowest By request:
concentration of DMSO (0.0078% v/v) ZIP/RAR file
Assay MCCL Form 4A -
after 72-hour incubation. of high-
Photo- Final Report [MTT] resolution
documen (APPENDIX) • Figure 3. A: <cell line> treated with photos in
-tation highest concentration of Doxorubicin JPEG/PNG
(X µg/mL) after 72-hour incubation. B:
<cell line> treated with lowest
concentration of Doxorubicin (Y µg/
mL) after 72-hour incubation.
• Figure 4. <cell line> treated with eight
concentrations (µg/mL) of <sample
name>

37
Service Deliverables & Final Reports

Table 5. cont. (3 of 4)
Service Deliverables and Final Reports, cont.
Other
Services Report Form Report Content Deliverables
• Methodology performed
• Table 1. Percent scratch areas (relative to Day
0) of <cell line> treated with the control
(DMSO)
• Table 2. Percent scratch areas (relative to
Day 0) of <cell line> treated with <sample
name> at X ug/ml. By request:
Wound • Table 3. Fold changes in wound closure ZIP/RAR file
healing/ MCCL Form 4B - (relative to the control) of <cell line> treated of high-
Final Report
Scratch [Scratch] with <sample name> at X ug/ml. resolution
Assay • Table 4. Pixel measurements of the scratch photos in
areas of treated <cell line>. JPEG/PNG
• Figure 1. A: Typical morphology of <cell line>
in T-25 culture flask before seeding. B: <cell
line> at 100% confluency prior to scratching.
• Figure 2. Scratched monolayers of <cell line>
treated with DMSO control and X µg/ml
<sample name> at Day 0, Day 1 and Day 3.

• Methodology performed
• Table 1. Percent Early Apoptotic, Late
Apoptotic and Necrotic cells of <cell line>
treated with control (DMSO)
By request:
• Table 2. Percent Early Apoptotic, Late ZIP/RAR file
Annexin MCCL Form 4C - Apoptotic and Necrotic cells of <cell line>
of high-
V-PI Final Report treated with with <sample name> at X ug/ml.
resolution
assay [Annexin V-PI] • Table 3. Summary of cell counts of 4 fields per photos in
treatment JPEG/PNG
• Figure 1. Representative fluorescence images
of <cell line> treated with control (DMSO) and
<sample name> at X ug/ml after 72-hr
incubation.

38
 Service Deliverables & Final Reports

Table 5. cont. (4 of 4)
Service Deliverables and Final Reports, cont.
Other
Services Report Form Report Content Deliverables

• Methodology performed
• Table 1. Summary of cell counts (6 fields)
and percent <marker>-positive <cell line>
treated with control (DMSO) By request:
ZIP/RAR file
IF
MCCL Form 4H - • Table 2. Summary of cell counts (6 fields) of high-
staining Final Report [IF] and percent <marker>-positive <cell line> resolution
assay treated with <sample name> at X ug/ml photos in
• Figure 1. Representative fluorescence JPEG/PNG
images of <cell line> treated with control
(DMSO) and <sample name> at X ug/ml
after 72-hr incubation.

39
G. Frequently Asked Questions

Does MCCL sell/donate cell lines?

No. Our cell lines are available because we are conducting ongoing research projects that
make use of these cell lines. We are not permitted by the original manufacturers (ATCC,
Lonza, Sigma) to resell these biomaterials. We only share/donate our cell lines to research
collaborators who are a part of our government-funded projects.

Does MCCL offer MTT assay for antimicrobial testing?

No. We only offer assays for phytochemical analysis and testing using mammalian (human/
mouse) cells.

Does MCCL offer plant/fungal identification services?

No. Please refer to the Institute of Biology website (https://biology.science.upd.edu.ph/) or e-
mail biology.upd@up.edu.ph to inquire about these services.

Does MCCL offer extraction services such as HPLC?

No. We do not offer any form of extraction service. Clients must perform their own crude
extraction/isolation protocols prior to submitting their samples to MCCL (See Section B)

Does MCCL offer testing/extraction of compound groups (eg. phytoestrols,

anthocyanins)?
No. We do not offer any form of extraction service. As for testing, we only offer analysis of
two major classes of phytochemicals: phenolics and flavonoids.

Does MCCL offer in vivo testing?

No. MCCL is primarily an in vitro research laboratory. We do not breed mice/hamsters/
zebrafish/brine shrimps for client services. In vivo testing requires ethical clearances that
MCCL cannot obtain for the client. For brine shrimp and chorioallantoic assays, please contact
andevlab.upd@up.edu.ph

Does MCCL provide technical consultations regarding the bioassays offered?

No. MCCL only functions as a service provider. The client must consult with their research
adviser/supervisor regarding data interpretations specific to their research.

Does MCCL provide trainings for clients?

Yes. MCCL offers cell culture training for registered clients once a year. Please refer to our
Facebook page (facebook.com/mccl.upd) or our website (https://sites.google.com/up.edu.ph/
mccl-upd-extension-services) for training announcements.

40
 Frequently Asked Questions

Can MCCL obtain a particular cell line specific for the client’s study?
No. Cell lines can cost around Php 50-200,000. Furthermore, cell line procurement is a long
process (around 6 months to 1 year). Hence, we only offer testing in cell lines that are already
available at our facility.

Can clients request customized sample concentrations for testing?

For Regular Assays (MTT, DPPH, TPC, TFC), these concentrations are fixed and non-
negotiable. For Confirmatory Bioassays (Annexin V-PI and IF staining), the specific sample
concentration/s are to be provided by the client.

Can clients request a different positive control for testing?

Yes. But the client must submit their preferred positive control and MCCL will treat it as a
submitted sample with the corresponding service fees.

Can clients come to MCCL and perform the assay protocols?

No. Our bioassay protocols may only be performed by MCCL personnel who have been
trained in performing these assays using highly sensitive equipment for at least 1 year. We
only provide this training for employed researchers and IB-enrolled thesis students, hence
clients without this training cannot be allowed to perform the assay protocols at MCCL.

Can clients come to MCCL and observe the assay protocols?

No. MCCL houses several researchers and research students who are involved under
government-funded research projects. Unfortunately, we cannot cater to visiting clients in
the interest of facility space and our busy research schedules.

Can clients perform their thesis/dissertation at MCCL?

Only graduate students enrolled at UP are being accepted to perform their dissertations at
MCCL even without official research collaboration.

Can MCCL sign my ISEF form?

No. The form can only be signed by lab workers/researchers who supervised the client’s
training and performance of experiments. Since the client is not allowed to perform their
assays at MCCL, our personnel cannot sign the ISEF form.

41
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Chen, J. (2016) The Cell-Cycle Arrest and Apoptotic Functions of p53 in Tumor Initiation and
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Crowley, L.C., Marfell, B.J., Scott, A.P. & Waterhouse, N.J. (2016) Quantitation of Apoptosis and
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 ACKNOWLEDGEMENTS

The MCCL bioassay services would not be possible without

the generous support of the following institutions:

43