1. Introduction to Cell Culture
Biotechnology I

2. Cell Culture
Definition: the in vitro growth of cells isolated from multi-cellular organisms
Process: Cells will continue dividing until they fill up the container; cell to cell contact stops cell division
Uses: vaccines, research of all kinds including stem cell, recombinant DNA, production of antibodies

3. Types of Cell used
Mammalian cells are commonly used but they do grow slower and are more fragile than bacterial cells. Also have more complex nutrient requirements
Bacterial cells were first used to make genetically engineered products and they reproduce very quickly

4. Growing Bacterial Cells
Choose bacteria – E. coli most common
Make media
Petri plates use agar in media (Luria Broth, nutrient agar)
Liquid cultures use broth (LB, nutrient broth)
Sterilize media in autoclave
Pour media plates
Innoculate media
Grow cells in incubator (37oC)

5. Equipment Required for Mammalian Cells
CO2 incubator: for proper atmospheric conditions
Laminar flow hood: for sterility
Hemocytometer: for cell counting
Inverted Microscope: for viewing cells on hemocytometer

6. Cell Culture Process Contaminating the cell culture is area of concern
To avoid contamination mfg. techs do the following:
Sterilize all containers used
Change the media frequently
Media is the source of nutrition, pH indicator often times included in media to help monitor pH which is critical to cell survival

7. Cell Culture Process Count cells
Using a hemocytometer the cells are counted to determine if they are ready to harvest
Can stain cells with trypan blue to determine if they are still alive. All cells which exclude the dye are viable. All stained cells are dead.
Live cell
Dead cell

8. Cell Culture Process View cells
Inverted microscope is used to determine cell growth and activity
Adherent cells are ones which are stuck to the flask or vessel
Cell counting video for cell culture