Test 2. Lab 7

Question 1

*What are terms used in cell passaging and define:

Answer 1

Cell seeding: transfer of a predetermined concentration of cells to a new vessel

Cell passaging: process of transferring cells to new vessels providing more media and space
It’s the passage of cells when they reach to 80-90% confluency in flask/dishes/plates

Confluencerefers to the proportion of the surface which is covered by cells.
Cells are typically passaged before becoming fully confluent in order to maintain their proliferation

Question 2

What are the steps of cell passaging?

Answer 2

1. remove old media
2. add PBS to wash cells

3 Add tripson that acts as a protease to remove cells from surface of plate

4. incubate
5. Remove plate from incubator and add media.
6. flush surface to mix cells in solution
7. add whole solution to new and sterile tubes
8. centrifuge solution
9. remove all media remaining cell pellet.
10. Add new media and break apart pellet by drawing and releasing media against wall
11. perform cell count using hemocytometer

Question 3

What is PBS solution?

Answer 3

a salt solution used to wash cells

Question 4

What is trypsin?

Answer 4

protease used to remove cells from the surface of the plate