1/11/2022

Aseptic Culture
Environment &
Mammalian Cell
Culture Lab
Equipment

Ankur Gautam
Lab Services Manager

Contents….

➢ Cell culture Environment

➢ Growth medium and its constituents

➢ Animal cell culture lab

➢ Cell culture equipment

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Animal Cell Culture Environment

Cell Culture Medium Nutrients

Supplements
Fetal Bovine serum

Sterile Environment No Microbes

Temp,
Controlled Environment Humidity
CO2

What is Important to Make Delicious Briyani?

Washing Nutrition Optimum Heat

Briyani

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What is Important to Grow Cells Healthy?

 Remove Unwanted  Retain Desired

Antibiotics, Growth
factors. Supplements

https://www.clipartguide.com/

Cell Culture Medium

Energy Source

Inorganic salts Buffering agents

Amino acids Vitamins

Most Commonly Used Commercial Media

➢ Dulbecco’s Modifies Eagle Medium (DMEM)

➢ Roswell Park Memorial Institute-1640 (RPMI-1640)
➢ Ham’s F12 Nutrient Mixture (f12)

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Fetal Bovine Serum

Provide growth factors & supplements

Growth Factors

Hormones
Macromolecules

Complete Medium

Cell Culture Medium

Bicarbonate system
• CO2 is required
• Economical
Energy Source
• Not toxic

Inorganic salts Buffering agents

Amino acids Vitamins

HEPES
• CO2 is not required
• Relatively expensive
• Prone to oxidation in light

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Bicarbonate System

❑ Natural buffering system

❑ Blood pH is maintained by this system

Mimicking the
in vivo
conditions

Weak acid Conjugate base

Air

Medium

Bicarbonate System

https://www.phe-culturecollections.org.uk/

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pH Changes Monitoring
Monitoring pH is very important (Twice in a day)

Change in medium pH- adversely affect the growth

Phenol Red is used as pH indicator

Can visually monitor the pH

https://www.phe-culturecollections.org.uk/

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Color Monitoring - Interpretation

https://www.phe-culturecollections.org.uk/

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NaHCO3 Vs CO2 concentration

CO2 Level

pH Value
https://blog.binder-world.com/en/adherent-cell-cultures

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NaHCO3 Vs CO2 Levels Physiologic pH

7.2 – 7.4
DMEM Medium
• NaHCO3 - 44mM
• CO2 requirement- 7.5% and 11%

EMEM Medium
• NaHCO3 - 26mM
• CO2 requirement- 4.5% and 6.5%

https://www.phe-culturecollections.org.uk/

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Cell Culture Medium Preparation

10%
50-100IU/ml penicillin
50-100µg/ml streptomycin.
FBS

Complete Medium

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Controlled Environment

https://www.benchmarkscientific.com/

https://www.labmanager.com/

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Microbe Free Conditions

https://www.thehindu.com/sci-tech/health/what-you-should-know-about-antimicrobial-
resistance-public-health-problem/article25652895.ece

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Biosafety Cabinet (level-II)

➢ Aseptic culture environments

➢ Unidirectional flow of HEPA-filtered air
➢ Prevents airborne contaminants

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CO2 Incubator

➢ Sterile environment with controlled parameters

➢ Mimicking in vivo conditions

https://www.benchmarkscientific.com/ https://devcell.bio.uci.edu/

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Inverted Microscope

To visualize the cell morphology

https://mfimedical.com/products/unico-iv953-inverted-microscope

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Centrifuge and Water bath

➢ To spin down cells

➢ To incubate medium and other reagents

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Liquid Nitrogen Storage Tank

➢ Storage of cell lines

https://www.thermofisher.com/

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Culture ware

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THANK YOU
ankur.gautam@merckgroup.com
https://www.linkedin.com/groups/10544590/

24 Chart library functional cells

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