Cell models

Question 1

What settings should usually the incubator be set at when culturing eukaryote cells?

Answer 1

37 degrees
5% CO2
Humidity of 85-95%

Question 2

What does a culture medium usually contain and what are the purpose of those contents?

Answer 2

• buffering system: stable pH
• phenol red (pH indicator)
• L-glutamine: It is an alternative energy source for rapidly dividing cells and cells that use glucose inefficiently
• Antibiotics: no contamination
• serum and/or other supplements and growth factors

Question 3

Name some reasons why cell models can be a useful tool?

Answer 3

• they can be designed to answer a specific question
• They can be very simple or rather complex
• The experiments can be repeated many times in a controlled way
• Single cells can be visualized and followed over time
• In the cell culture dish there is possible to speed up the pathological processes
• time and cost effective

Question 4

What type of cells can a cell culture model be based on?

Answer 4

• cell lines
• primary brain cells from research animals
• stem cells from research animals
• human brain cells derived from ES (embryonic stem cells) or iPS cells

Question 5

What are the advantages/disadvantages with cell lines

Answer 5

Advantage: 
- easy to expand 
- cheap 
easy to transfect 
- no ethical permission needed
- human cells available 

Disadvantage:
-tumor cells are not normal cells

Question 6

What kind of research are cell lines suitable for?

Answer 6

studies of basic cellular mechanisms that are general for most cells

Question 7

What are the advantages/disadvantages with primary brain cells from research animals

Answer 7

Advantages:

• specialized cells can be isolated
• well established protocols

Disadvantage::

• ethical permission needed
• differences between human cells and animal cells
• expensive

Question 8

What kind of research are primary brain cells from research animals suitable for?

Answer 8

studies of mature neurons or glial cells

Question 9

What are the advantages/disadvantages with stem cells from research animals

Answer 9

Advantages:

• the cells can be expanded which means less cost and fewer animals are needed
• well established protocols
• neural stem cells can be differentiated to neurons and glia

Disadvantages:

• ethical permission needed
• differences between human cells and animal cells
• the neurons in the culture will be immature

Question 10

What are the advantages/disadvantages with stem cells from research animals

Answer 10

advantages:
- can be expanded and differentiated to various cell types
- enables studies of human specialized cells and studies of cells from patients with a specific disease

Disadvantages:

• expensive
• differentiation takes long time and can result in immature cells

Question 11

What kind of research are human brain cells derived from iPSCs suited for?

Answer 11

studies of human brain cells

Question 12

Name 4 types of 3D culture models

Answer 12

• microfluidic organs-on chips
• Organotypic brain slices
• neurospheroids
• cerebral organoids

Question 13

What are microfluidic organs-on chips?

Answer 13

created by culturing several layers of living cells in a perfused chambers of a microfluidic cell culture device to resemble organ level physiology

Question 14

What are Organotypic brain slices?

Answer 14

created by culturing tissue slices from whole postnatal or adult brains to resemble the in vivo conditions of high density cells

Question 15

What are neurospheroids?

Answer 15

created from single suspensions of neural stem and progenitor cells derived from adult or fetal CNS as well as from embryonic stem cells

Question 16

What are cerebral organoids?

Answer 16

created by culturing human pluripotent stem cells in a three dimensional rotational bioreactor over a course of months

Question 17

What are the advantages/disadvantages with organoids?

Answer 17

advantage:
- human organoids can mimic the human organ and are more relevant for predicting drug toxicity than animal models

Disadvantage:

• difficult and expensive models to work with
• Organoids may only recapitulate the first few months of development
• do not mimic an organ compleatly (e.g. lacks support scaffolds, blood vessels and interactions with other organs of the body)

Question 18

Name ways to induce AD or PD pathology in cell culture model systems

Answer 18

• overexpress pathological A-beta, tau or alpha synuclein
• add A-beta, tau or alpha synuclein to the cell culture medium
• use mutated cells from transgenic mice or iPS cells from patients
• use viruses or CRISPR technology to induce mutations