BIO265 Cell Culture Techniques

In vivo

research performed on living organism

What is a con of in vivo research?

can only obtain measurement at one time point

In vitro

• “in glass”, research performed under controlled conditions outside of organism

• often involves cell/tissue culture

What are pros and cons of in vitro research?

• pros: more control over system, more dynamic information from multiple time points, clear and more detailed imaging

• cons: artificial environment so cannot replicate all natural responses to stimuli

What is cell culture?

growth and maintenance of cells outside of a living organism (in an artificial environment)

What are some examples of cell culture?

• immortalized cells (HEK-293, HeLa) - easy

• stem cells (natural proliferation)

• primary culture (e.g., explant tissue)

• EACH HAS DIFFERENT DEMANDS

What are some pros to cell culture?

• researchers can manipulate culture environment/conditions (w/ culture media)

• less expensive (most of the time)

• faster than many in vivo studies (these require many assumptions & precise technique)

What are some limitations to cell culture?

• Are immortalized cells a good model? → they are under different selective pressures

• Do cells in a dish behave similarly to cells in an organism? → not quite, but there are similarities

What are examples of cell culture equipment and reagents?

• biosafety hood (protects cells from contamination)

• cell incubator (37C for humans cells→ matches internal body temp)

• cell culture flasks/plates

• water bath (warms up media before it is added to cell culture)

• microscope (visualization)

• centrifuge

• fridge/freezer

What does cell culture media/growth media do?

provides nutrients, glucose, and growth factors necessary to promote survival and proliferation

What is cell culture media composed of?

• basal media

• serum

• antibiotics

What is basal media?

• contains amino acids, vitamins, glucose, and buffering agents (sodium bicarbonate) to maintain physiological pH (7.4) in a 5-10% CO2 environment →

• e.g., Dulbecco’s Modified Eagle Media (DMEM)

• meets minimum needs and composition depends on cell type

What is serum?

• often Fetal Bovine Serum or FBS (because cells are still growing & developing)

• provides growth factors necessary for survival and proliferation

What are antibiotics used for in cell culture media?

• prevent growth of bacteria

• may also be used for selection