Tissue Culture

Question 1

What cells did we use in tissue culture?

Answer 1

Cos-7 monkey KIDNEY cells

–> Fibroblast like cells derived from monkey-kidney tissue

Question 2

What are some features of Cos-7 cells that make them ideal for tissue culture?

Answer 2

• fast growing
• adherent
• Easy to transfect
• capable of 100% confluency

Question 3

What is confluency?

Answer 3

Confluency refers to the % of a cell surface that is covered by adherent cells (e.g. 100% refers to a cell surface that is completely covered in a monolayer)

Question 4

The media used for tissue culture was DMEM. What are the components of DMEM?

Answer 4

• Nutrients, growth factors required for cell growth (e.g. folic acid)
• 10% Foetal Bovine Serum: is deprived of cells, fibrin and clotting factors but contains factors essential for cell growth
• 1% Penstreptomycin (penicillin and streptomycin)
• Trypsin inhibitor (to stop trypsinastion process)
• Phenol red dye (acts as a pH indicator. Pink-red is healthy (pH=7); yellow indicates accumulation of waste material and cell death, so medium needs to be changed

Question 5

What is the function of PBS?

Answer 5

Cell-rinsing and remove media

contains inhibitors that stop trypsin from working

Question 6

What is the function of trypsin EDTA

Answer 6

Trypsin is a serine protease that detaches cells and prevents cell-cell adhesion
Allows for resuspension of the cells as well during splitting
EDTA is a metal chelator (Ca2+; mg2+); enhances trypsin activity