lecture 4 - pluripotent stem cells II

Question 1

What are the different mouse ESC states?

Answer 1

Naive ground state, intermediate state, primed state - can then differentiate into ectoderm, endoderm and mesoderm

Question 2

How can hESCs be rewired to a naive state?

Answer 2

Is the naive pluripotent state of hESCS inherently unstable in current conditions used?
Used ectopic induction of OCT4, KLF2 and KLF4 expression
Cultured in LIF, plus GSK-3 and MEK inhibitors
These converted hESCs properties similar to naive mESCs
they therefore generated naive hESCs in a stabilised pluripotent state

Question 3

What four transcription factors can be used to reprogram mouse fibroblasts to pluripotency?

Answer 3

Oct-4, Sox-2, C-myc and Klf-4

Question 4

What are some potential applications of iPSCs?

Answer 4

Cell replacement therapies - regenerative medicine
disease in a dish models
drug discovery
cellular agriculture

Question 5

What is an advantage of using iPSCs in drug discovery?

Answer 5

They can reduce attrition rates in drug discovery, which is a problem when using human hepatic cells

Question 6

How can CRISPR/Cas9 editing be used in hPSC cardiomyocytes?

Answer 6

Shows potential for the pharmacological rescue of arrhythmias, pointing towards new targets for mechanistic understanding and therapeutics

Question 7

What are some applications of using organdie cultures?

Answer 7

Modelling human disease - intestinal organdies from CF patients for drug discovery
Can be used in target identification, drug screening and toxicity testing

Question 8

How can stem cells be used for livestock and food security?

Answer 8

Can obtain pluripotent embryonic stem cells from livestock and culture in the presence of active A, FGF and Wnt inhibition
this can be used for enhanced animal breeding and production of lab-grown meat from a renewable stem cell source

Question 9

What is an example of how HSCs can be used to model disease?

Answer 9

Viral diseases such as hepatitis - provide a platform for studying mechanisms of infection and the life cycle of a virus or parasite
however, these current models are not suitable for modelling drug toxicity or injury, so protocols for their generation need to be improved

Question 10

How can patient-derived hiPSCs be used to study genetic disorders?

Answer 10

• For example, systemic lupus erythematosus (SLE) is a kind of chronic inflammatory autoimmune disease that can impact any part of the body
• The pathogenesis involves dysfunction of both specific and non-specific immunity, but the aetiology remains unclear – no effective cure
• Current immunosuppressive therapy is of high cost, with serious side effects
• The phenotype and stage of progression are various, so personalised treatment is needed for a better effect
• By using mononuclear cells of SLE patients, Li et al generated SLE patient-derived hiPSCs successfully, which offers a tool for exploration of the SLE disease mechanism and drug discovery