Bio 363 Human Genetics Spring 2013
Stem Cells study sheet
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Background:
What organs have Dr. Atala and his colleagues worked on regenerating? What are the general approaches? How do stem cells play into these efforts?
What are the different types of stem cells that are naturally occurring?
What is therapeutic cloning?
What are iPS cells?

Hipp et al, 2010:
How would you describe, in general, the gene expression profile of stem cells compared to differentiated cells?
What cited finding is consistent with the hypothesis that expression of differentiation genes does not necessarily reflect partial differentiation?
What cited finding suggests that transdifferentiation is possible?

Tian et al., 2010:
How common are mesenchymal stem cells among all bone marrow stromal cells?
Into what cell types can MSCs from the bone marrow differentiate?
What is conditioned medium?
What are the two approaches for getting BMSCs to differentiate for tissue engineering?
What were the three goals of this study?
Fig 1: Does coculture give the same result as use of conditioned medium? What negative control is apparently not shown but mentioned in the text?
Fig 2: What’s the difference between the genes examined in A vs. B? Do the gene expression results from coculture vs. conditioned medium reflect what we saw in the previous figure?
Fig 3: Is there any gene/gene product showing inconsistent or puzzling results here compared to Fig 2?
Fig 4: What type of induction is used? What are the controls? Which color represents the gene product in question? What does the other color show? Which gene products are present without induction? What is consistent or not with previous figures?
Fig 5: What type of induction is used? What do colors represent? Are results consistent with previous figures?
Fig 6: What technique was used to get these data? What growth factors are present under different conditions?
Fig 7: What are the logistics of this experiment? What’s the best way to regenerate truly contractile tissue?
Fig 8: What are the logistics? What feature of which panels indicates muscle tissue formation?

Zhao et al., 2012
What are the main dangers of using iPS cells?
What sorts of strategies have been used before to address this, and and what were the worries with these strategies themselves?
What general strategy is used here?
What is the novelty? (see beginning of discussion)
What is episomal DNA, and what is unusual about the S/MAR sequence’s effect on episomal DNA in human cells?
Fig 1: They start with a proof of concept experiment that uses GFP as a readout. What are the main features of the plasmid in panel C? How should it behave after transfection? How do the data in panels D-F fit together?
Fig 3: How is this construct different from the previous one? What promoter drives TK, and what does TK make cells sensitive to? What are NCCIT cells? Note that RA is retinoic acid, which drives differentiation of NCCIT cells. OCT4 is a protein expressed in stem cells but not differentiated cells.
Panel C: can you transform NCCIT cells with this construct and have the construct maintained?
Panel D: This is a key result! What is blue and what is red? What does the construct enable?
Panel E: What is on the x axis? What do the peaks represent? How does this fluorescence activated cell sorter (FACS) experiment confirm the results in panel D?
Fig 5: What are the logistics of this experiment? Can this approach help control cancer formation from implanted cells?