GROUP 9

CELL CULTURE
Members:

Fernandez Hernaly, Lanticsi Faye Marie, Sacares Joshua

Daryl, Tampus Princes Alicia / BS BIOLOGY 3A

WHAT IS CELL
CULTURE?

Cell culture refers to the removal of cells from an

animal or plant and their subsequent growth in a

favorable artificial environment.

The cells may be removed from the tissue directly

and disaggregated by enzymatic or mechanical

means before cultivation, or they may be derived

from a cell line or cell strain that has already been

established
 HISTORY OF CELL
CULTURE
Sydney Ringer, an English physiologist from the Cell culture techniques progressed considerably in the

nineteenth century, produced salt solutions 1940s and 1950s to help virology research. Growing

comprising sodium, potassium, calcium, and viruses in cell cultures allows for the purification of

magnesium chlorides that could keep an isolated

viruses for the production of vaccines. Jonas Salk's

animal heart beating outside the body. Wilhelm

injectable polio vaccine was one of the first items mass-
Roux established the idea of tissue culture in 1885
produced utilizing cell culture techniques. The cell
when he took a piece of an embryonic chicken's
culture research of John Franklin Enders, Thomas Huckle
medullary plate and kept it in a warm saline
Weller, and Frederick Chapman Robbins, who were given
solution for many days. From 1907 to 1910, Ross
the Nobel Prize for discovering a technique of cultivating
Granville Harrison, working at Johns Hopkins
the virus in monkey kidney cell cultures, made this
Medical School and subsequently at Yale
vaccination conceivable.
University, reported the findings of his research,

developing the methodology of tissue culture.

TIMELINE OF THE HISTORY OF CELL CULTURE
 PRIMARY CELL CULTURE SECONDARY CELL CULTURE

Derived directly from excised tissue and cultured either When a primary culture is sub-cultured, it

as: becomes secondary culture or cell line.

Outgrowth of excised tissue in culture Subculture (or passage) refers to the transfer

Dissociation into single cells (by enzymatic of cells from one culture vessel to another

digestion or mechanical dispersion) culture vessel.

Advantages: Derived from a primary cell culture.

usually retain many of the differentiated Isolated by selection or cloning.

characteristics of the cell in vivo Becoming a more homogeneous cell

Disadvantages: population.

initially heterogeneous but later become Finite life span in vitro.

dominated by fibroblasts. Retain differentiated phenotype.

the preparation of primary cultures is labor

Mainly anchorage dependant.

intensive • can be maintained in vitro only for a

Exhibit contact inhibition.

limited period of time.

CONTINUOUS CELL LINE CELL STRAIN
CELL LINES
A cell strain is created by selecting or
Continuous cell lines are generated A cell line is a permanently formed cell

cloning cells with specified features

when cell lines are transformed in a culture that will grow forever if provided

with adequate fresh media and space. or characteristics that must be

laboratory or in vitro growing
Lines are distinguished from cell strains in determined from a parent culture or
settings. The cell lines exhibit ploidy
that they have surpassed the Hayflick a cell line. Cell strains are culture-
(aneupliody(abnormal chr no)) in the
limit and become immortal. Some
adapted cells that, unlike cell lines,
absence of contact inhibition and
animals, notably rodents, produce lines
have a restricted division potential.
anchorage dependency. They can
very easily, whereas others do not. There
Non-immortalized cells cease to
develop as a monolayer or as a have been no cell lines created from
divide after 40 to 60 population
suspension. The pace of growth is avian tissues, and creating cell lines from
doublings and lose their capacity to
fast, with a doubling period of 12-24 human tissue is challenging. Many cell

proliferate (a genetically determined

hours. biologists believe that a cell line is

already aberrant and is on its path to event known as senescence).

becoming the culture equivalent of a

neoplastic cell.
GROWTH GROWTH CONDITIONS

CONDITIONS, Growth medium

Controlled temperature

ADVANTAGES CO2

AND Relative Humidity

Substrate for cell attachment

DISADVANTAGES Incubator that maintains correct pH and osmolality

OF CELL CULTURE
ADVANTAGES

In the in vitro system, the physio-chemical environment in the culture

can be precisely controlled.

Physiological conditions in the cell culture can be controlled.

The use of selective media in cell culture techniques allows us to

maintain cell homogeneity.

Using cytological or immune-staining techniques, cells in culture

can be easily identified.

With a suitable cryopreservation medium such as DMSO, cell

cultures can be stored in liquid nitrogen for a very long time.

GROWTH
DISADVANTAGES
CONDITIONS, The most difficult part of cell culture is maintaining a sterile

ADVANTAGES aseptic condition.

AND The in vitro methods have a high risk of chemical and

microbial contamination.

DISADVANTAGES Different types of cells in culture have a high risk of cross-

OF CELL CULTURE contamination.

For most cells, experience and expertise are required for

effective maintenance.

Requires a thorough standardization of the medium, nutrient

concentration, and serum. All of this varies depending on the

type and origin of the cells.

WHAT IS
APOPTOSIS?
Apoptosis is a form of programmed cell

death or pre-planned cell death.

It is important to maintain cellular

balance.

It is used during early development to

eliminate unwanted cells.

 During apoptosis, the condensation of

chromosomes takes place.

membrane blebbing will take place

during apoptosis, which will result in the

destruction of the cell.

Mechanism
Intrinsic Pathway or Mitochondrial

Pathway
 WHAT IS SENESCENCE?

Senescence is the deterioration that takes place

following the concept of ageing.

During senescence, the cell cycle is inhibited or

blocked at different entry points of the cycle.

Genetics play a major role in senescence.

Cellular senescence has been identified at specific

developmental windows and at discrete sites during

embryo development

Senescence canact as a potent anti-tumor

mechanism
 WHAT IS THE DIFFERENCE
WHAT ARE THE SIMILARITIES BETWEEN APOPTOSIS AND
BETWEEN APOPTOSIS AND SENESCENCE?
SENESCENCE?
APOPTOSIS IS PROGRAMMED
APOPTOSIS AND SENESCENCE
CELL DEATH, WHICH IS PRE-
ARE TWO PROCESSES THAT
DETERMINED, WHILE
CAUSE CELL DEATH.
SENESCENCE HAPPENS OVER
THEY ARE COMPLEX
AGEING AND IS NOT PRE-
PROCESSES WITH VARIED
DETERMINED.
MECHANISMS.
PROTEOLYTIC MECHANISMS
GENETICS PLAY A MAJOR ROLE
PLAY A MAJOR ROLE IN
IN BOTH APOPTOSIS AND

SENESCENCE. APOPTOSIS, WHEREAS GENETIC

MECHANISMS PLAY A MAJOR

ROLE IN SENESCENCE.
WHAT IS CELL Cell morphology pertains to the morphological features

of the cell(s). It employs microscopy to identify the

MORPHOLOGY? shape, structure, form, color, texture, pattern, and size

of the cell(s). In bacteriology, for instance, cell

morphology pertains to the shape of bacteria if cocci,

bacilli, spiral, etc., and the size of bacteria. Thus,

determining cell morphology is essential in bacterial

taxonomy.

Synonyms: cellular morphology; cytomorphology.

TYPES OF BACTERIAL
CELL MORPHOLOGY

Coccus (plural cocci):

Spiral: twisted like a DNA

Spherical shape, like tiny

helix

balls

Bacillus (plural bacilli): Rod

Vibrio: comma-shaped
shape, like cylinders
 Cell Morphology Examples of
Prokaryotes

1. Staphylococcus spp., Streptococcus

DIFFERENT CELL spp., Micrococcus spp., Neisseria

MORPHOLOGIES spp., Gonococcus, Sarcina spp.,

1. Coccus
IN PROKARYOTES Moraxella spp., Enterococcus spp.

2. Bacillus spp., Clostridium spp.,

2. Bacillus
Escherichia spp., Klebsiella spp.,

3. Spiral Pseudomonas spp., Proteus spp.,

Salmonella spp., Actinomyces spp.

4. Vibrio
3. Spirillum spp., Campylobacter spp.,

Helicobacter spp., Leptospira spp.,

Borrelia spp., Treponema spp.

4. Vibrio cholerae, Vibrio

parahaemolyticus, Vibrio vulnificus,

Vibrio gastroenteritis, Vibrio harveyi

(A) Neisseria gonorrhoeae or the Gonococcus is a gram-negative

coccus-shaped bacterium. It causes gonorrhea, a sexually-

transmitted disease. Credit: CDC/ Joe Millar.

(B) Streptococcus pneumoniae is a gram-positive coccus-shaped

bacterium. It is one of the main causal organisms of pneumonia.

Credit: CDC/ Dr. M.S. Mitchell.

(C) Staphylococcus aureus is a gram-positive coccus-shaped

bacterium. It, along with Streptococcus pyogenes, causes the

toxic shock syndrome. Credit: CDC/ Matthew J. Arduino, DRPH.

(D) Escherichia coli, the model organism used for bacterial

studies is a rod-shaped bacterium. Credit: Eric Erbe/USDA.

(E) Treponema pallidum is a spiral bacterium that causes syphilis.

Credit: CDC/ Dr. David Cox.

(F) Helicobacter pylori is a spiral bacterium that causes peptic

ulcers by damaging the duodenum in the digestive system. Credit:

John Hopkins Medicine.

(G) Vibrio cholerae causes cholera- an intestinal infection.

Source: Denniss, public domain.

Eukaryotes are organisms that have a well-defined nucleus

and specialized organelles in their cells. They can be both

unicellular or multicellular. Hence, it’s not necessary that the

CELL
cell morphology of eukaryotic cells will define the eukaryotic

organism’s morphology.

In the medical context, cell morphology is the phenotypic MORPHOLOGY

presentation of a cell tightly linked with its role in a

eukaryote’s body. In eukaryotes, mostly all organisms are

EXAMPLES IN
multicellular. The cells are in synchrony with each other to

make the body functional. Hence, a hierarchical system is as

EUKARYOTES
follows: cells » tissues » organs. Different tissues of
eukaryotes possess different types of cells based on varying

morphologies.
TYPES OF CELL
MORPHOLOGY IN
EUKARYOTES
PLANT CELL MORPHOLOGY: PLANT CELLS HAVE A

DISTINCT CELL WALL WHICH GIVES PLANT CELLS A

HIGHLY DEFINED MORPHOLOGY. THE SHAPE

VARIES FROM TISSUE TO TISSUE. EPIDERMAL

CELLS ARE GENERALLY RECTANGULAR WHILE

CORTEX CELLS ARE ISODIAMETRIC. SOME

SCLERENCHYMA CELLS ARE STONE-SHAPED

WHILE PARENCHYMA CELLS RANGE FROM NEARLY

SPHERICAL TO BRICK-LIKE SHAPE.

 Mammalian Cell Morphology:
Mammalian cells are basically of 3 types:

Within these 3 major

types, there are

further sub-types. Like

epithelial-like cells

b) Fibroblast-like cells: can be squamous,

c.) Lymphoblast-like
a) Epithelial-like cells: columnar, or cuboidal.
Spindle shaped and flat,
cells: Spherical, round
Rectangular shaped,
either bi- or multi-polar

shaped, can grow in

kind of regular
{Both epithelial-like and
suspension (no need to
dimensions
fibroblast-like cells attach
attach to a surface!!)
to a surface and grow!!}

 Figure 2: Fibroblast-
Figure 1: Synovial fibroblasts
like cells

Figure 3: Figure 4: We observe fibroblast- Figure 5:

Epithelial-like cells like cells in people affected with Lymphoblast-like cells
RA (rheumatoid arthritis).
 Interesting Fact!

Cell morphology is an exceptional field of Science that can help

you in identifying when the cells of your body start behaving

“abnormally”!!!

YES, just by closely observing a cell’s morphological characteristics

like its shape, nucleus size, cytoplasmic content, invaginations,

attachment to basal membrane, etc, we can tell if it is healthy or

not. When a tumor cell grows in your body, it can be easily be

distinguished because of the altered morphology- irregular shape,

nucleus and nucleoli become bigger than normal, cytoplasm seems

depleted, change in color of cytoplasm, detachment from the basal

membrane in case of malignant tumors. These changes are

indicative of cancerous growth! Also, for invasiveness and

metastasizing, malignant tumors start producing some “lytic factors”

which upon release, eat up the basal membrane and set the

malignant cancerous cells “loose”.

Modern methods of in-vitro single cell-derived clones “MORPHO-

PROFILING” can provide us probable avenues and possibilities to

identify cancerous and metastatic potentials of cells. This is how

advancing Science paves way for precision medicines!!! Source:

(Wu, 2020)
Animal Cell Culture: Concept map

FIGURE 7: ANIMAL CELL CULTURE

 Signs of deterioration of cells include
Regularly examining the morphology of
granularity around the nucleus,
the cells in culture (i.e., their shape
detachment of the cells from the
and appearance) is essential for
substrate, and cytoplasmic vacuolation.
successful cell culture experiments. In
Signs of deterioration may be caused by
addition to confirming the healthy
a variety of reasons, including
status of your cells, inspecting the cells
contamination of the culture, senescence
by eye and a microscope each time
of the cell line, or the presence of toxic
they are handled will allow you to
substances in the medium, or they may
detect any signs of contamination
simply imply that the culture needs a
early on and to contain it before it
medium change. Allowing the
spreads to other cultures around the
deterioration to progress too far will
laboratory.
make it irreversible.
FIGURE 8:
GENERAL
CONCEPT
MAP OF
THE CELL
CULTURE
THANK YOU
ANY QUESTION?