Lesson 1 - Intro to Cell Culture and Cell Types

Question 1

What is the ultimate goal of Cell biology?

Answer 1

To understand how macromolecular systems and organelles work and cooperate to enable cells to function autonomously and in tissues

Question 2

What do Eukaryotic cells contain?

Answer 2

A nucleus containing DNA

Question 3

What is Cell culture? How do we study it?

Answer 3

• the technique used to grow cells or tissues outside the organism under strictly controlled conditions outside of the body
• need to have reagents and equipment
• the cells of tissue of interest need to break apart

Question 4

What do glycoproteins have?

Answer 4

• sugar molecules associated with them to help stick to one another

Question 5

How are cells isolated form tissue? What are the steps?

Answer 5

• cells are isolated by breaking down cell - cell and cell - matrix interactions
• MECHANICAL METHODS –> taking a tissue and cutting it into smaller pieces, pushing through mesh to get smaller clumps of cells
• ENZYME –> chews up glycoproteins
• EDTA –> sponge that soaks up magnesium and calcium to help the cells dissociate

Question 6

What happens after cells are isolated?

Answer 6

• cells are supplied with proper nutrients in a liquid ,serum cells, and grown at optimal temperature

Question 7

How are serum cells grown?

Answer 7

• from cows
• liquid within the blood contains insulin which is a signaling molecule that allows cells to intake glucose and growth factors

Question 8

What is an incubator?

Answer 8

• mimics conditions found in the body
• tanks diffuse CO2 into the incubator which reacts with sodium bicarbonate to maintain the pH of the media
• phenol red indicated proper pH at 7.2
• cells adhere to the bottom of the agar plate

Question 9

How can cells be prevented from adhering to the bottom of the plate?

Answer 9

• by agitating the agar plate with various instruments

Question 10

What is a primary cell culture?

Answer 10

• refers to cell taken directly from an organism, they usually divide a limited number of times and under contact inhibition of cell density is high

Question 11

What is the process within the primary cell culture? What are the limitations?

Answer 11

• add trypsin to the plate of primary cells which causes them to dissociate
• dilute the cells down to a low level and put them on a fresh plate so that they can adhere
• repeat process
• can only do this a limited number of times

Question 12

What is the hayflick limit?

Answer 12

• everytime a cell divides, telomeres shorten until they reach a critical point called cellular senescent
• they will not grow anymore

Question 13

What is the cell line?

Answer 13

• refers to cells which are transformed and able to grow indefinitely,
• known as immortal cells
• less likely to exhibit contact inhibition

Question 14

What are transformed cells?

Answer 14

• have some dysregulation of the cell cycle regulatory mechanism and divide indefinitely and do not exhibit contact inhibition

Question 15

What were the findings of a HeLa cell?

Answer 15

• the cancer cells doubled in 24 hours
• cancer cells were immortal
• made many contributions to medical research but the cells were taken without Henrietta’s permission

Question 16

Normal cells exhibit :

Answer 16

• elongated, aligned, parallel arrays, contact inhibition

- take the fiberglass and infect them with an oncogene to put them into a transformed state

Question 17

transformed cells exhibit :

Answer 17

• rounded, hair like processes, disorganized, don’t experience contact inhibition, cancer like state, grow indefinitely

Question 18

Why are transformed/primary cells of value?

Answer 18

• primary : we want to understand normal physiology, more closely in the physiological state of the organism
• transformed : much more easy to grow, predictable behaviour, adapted to culture, reproducible results