Mammalian Cell Culture Webinar Series 7/30/2020 Webinar #1 · Ankur Gautam, Ph.D. Scientist Merck...

Mammalian Cell Culture Webinar SeriesWebinar #1

7/30/2020

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Mammalian Cell Culture Webinar Series

Join our LinkedIn group for more updates

https://www.linkedin.com/groups/10544590/

Date Topic

27.07.2020 Introduction to Mammalian Cell Culture

10.08.2020 Introduction to Cell Culture - Best Practices and

Challenges

24.08.2020 Cell Culture in 3D synthesis - Moving from 2D to 3D

environment

Ankur Gautam, Ph.D.Scientist

Merck High-End Skill Development

Centre

27th July, 2020

Mammalian Cell Culture Webinar Series #1:

Introduction to

Mammalian Cell Culture

Contents….

➢ History

➢ Animal Cell Culture

➢ Importance & its Applications

➢ Types of Cells

➢ Morphologies of cells

➢ Cell Culture Environment

➢ Growth Medium and its Constituents

➢ Animal Cell Culture Lab

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19121940s

19551961

19651981

2006

Chicken heart muscle

tissueculture

Antibiotics in medium

DMEM

Isolated human

fibroblasts

Serum-free medium

Mouse Embryonic Stem Cell

Induced Pluripotent Stem Cell

Shinya Yamanaka

Nobel Prize in Medicine (2012)

History

What is Mammalian Cell Culture ?

“Culturing of cells derived from animal cells/tissue”

Growing animal cells outside of the body under controlled conditions

Animal Tissue Culture of cells

Artificial environment: Similar to in vivo conditions

Why Cell Culture ?

➢ Cell is the Basic Unit for the Life

➢ Excellent model systems

Don’t test on me, rather test on cells

Lets test my new drug on

you?

➢ Reduce the animal usage

https://www.123rf.com/https://www.vectorstock.com/

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Production of vaccine & therapeutics

Function & Biochemistry

Drug & toxicity

screening

Applications of Cell Culture

Dr. Jonas Edward Salk (1914-95)

Cell Types

Immortalize

Cells

Secondary

Cells

Primary

Cells

Primary Culture cells

▪ Limited life span

▪ Difficult to maintain in culture

▪ Slow proliferation

▪ Not well characterized

▪ Derived directly from living tissue

▪ More similar to in vivo studies,

▪ Clinical relevance;

▪ Translational research

Excision of

tissue

Enzymatic/

mechanical

treatment

Single cell

Separation

Culture

DisaggregationMince or chop

Heterogenous

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Secondary Culture Cells

▪ Derived from primary cells

▪ Isolated by selection

▪ More homogeneous

▪ Finite life span in vitro

Selective medium

Heterogenous

Cell Type A Cell Type B

Homogenous

Primary culture

Secondary culture Secondary culture

• Modifications; transformed

• Altered physiological properties

• Not representative of in vivo

• Derived from pri / sec culture

• Immortalized, Fast growth

• Infinite life span

Continuous/Immortalized cells

Secondary cultureContinuous culture

Loss of cell division Control

Primary cells culture

(Heterogenous)

Secondary cells culture

(Homogenous)

Continuous cells culture

(Homogenous)

Definite time

Slow

Definite time

Slow

Indefinite time

Fast

Animal

Transformation

Selective medium

Disaggregation

Origin of Cell line

Tissue

Cell lines

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Types of Cells (Anchorage)

Anchorage dependant (Adherent)

▪ Most animal tissue derived cells

▪ Adhere to bottom of a flask

▪ Form a monolayer

Anchorage independent (Suspension)

▪ Cells associated with body fluid

(eg. blood cells)

▪ Free-floating (in suspension)

https://www.labconscious.com/

Fibroblast(Elongated & Bipolar)

Neuron(Long extension)

Epithelial(Flattened & Polygonal)

Lymphoblast(Spherical & suspension)

Types of cells (based on morphology)

Cell line Species of origin Tissue of origin Cell morphology

3T3 Mouse Connective Fibroblast

CHO Chinese Hamster Ovary Epithelial

BHK21 Syrian Hamster Kidney Fibroblast

HeLa Human Cervical Carcinoma Epithelial

MDCK Dog Kidney Epithelial

MRC-S Human Lung Fibroblast, Finite

Example of some cell lines used for in vitro culture

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Have You Ever Heard of The Immortal HeLa cells?

African lady, In USA

Cervical cancer (1951)

Surgical removal of tumor

Cells grown in laboratory

Ms. Lacks’ family didn’t know until 24 years after her death.

75 000 scientific article abstracts 11 000 patents.

First immortal Human cells grown outside the body.

Widely used cell line

Across the globe

Henrietta Lacks

https://www.gesundheitsindustrie-bw.de/en/

Movements of

continuous cells/

Cell lines

https://gfycat.com/joyfulgoldencoyote

https://www.shutterstock.com/

Animal Cell Culture Environment

Cell Culture Medium Nutrients

Fetal Bovine serum Supplements

Sterile Environment No Microbes

Controlled Environment

Temp,

Humidity

CO2

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Washing

What is Important to Make Delicious Briyani?

Briyani

Nutrition Optimum Heat

Remove Unwanted Retain Desired

Antibiotics, Growth factors. Supplements

What is Important to Grow Cells healthy?

https://www.clipartguide.com/

Cell Culture Medium

Amino acids

Inorganic salts

Vitamins

Buffering agents

Energy Source

➢ Dulbecco’s Modifies Eagle Medium (DMEM)

➢ Roswell Park Memorial Institute-1640 (RPMI-1640)

➢ Ham’s F12 Nutrient Mixture (f12)

pH 7.2 to 7.4

Most Commonly Used Commercial Media

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Fetal Bovine Serum

Provide growth factors & supplements

Hormones

Growth Factors

Macromolecules

Complete Medium

Controlled Environment

https://www.benchmarkscientific.com/

https://www.labmanager.com/

Microbe Free Conditions

https://handling-solutions.eppendorf.com/

https://www.dreamstime.com/

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

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Animal Cell Culture Laboratory

Biosafety Cabinet (level-II)

➢ Aseptic culture environments

➢ Unidirectional flow of HEPA-filtered air

➢ Prevents airborne contaminants

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

Centrifuge and Water bath

➢ To spin down cells

➢ To incubate medium and other reagents

Liquid Nitrogen Storage Tank

➢ Storage of cell lines

https://www.thermofisher.com/

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Cultureware

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[email protected]

[email protected]

[email protected]

[email protected]

Our Team

[email protected]

[email protected]

[email protected]


New students please send their details

THANK YOU

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mailto:[email protected]

mailto:[email protected]



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Mammalian Cell Culture Webinar Series

Join our LinkedIn group for more updates


Date Topic

27.07.2020 Introduction to Mammalian Cell Culture

10.08.2020 Introduction to Cell Culture - Best

Practices and Challenges

24.08.2020 Cell Culture in 3D synthesis - Moving from 2D to 3D

environment

34


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