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2 - Krause_hili

2 - Krause_hili - transplantation experiment How did the...

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Krause paper Highlights What is meant by LTR, . ..STR, . ..... homing? Define a niche What is the HSC niche? How does this relate to homing? Understand the homing purification protocol. Why do they need to go through all this? How does elutriation work? What is lineage depletion? and how is it performed? How are the cells labeled and how does this allow the homed cells to be recovered? Where does FACS come into it? What is required for survival of a bone marrow transplant? answer: the animals are injected with a population of STR cells and must get at least one LTR HSC cell to survive What is FISH and how was it used to identify donor cells DNA probe allows detection of DNA by hybridization Probe is biotinylated, and a fluor attached to avidin is used to visualize it. Can FISH and immunofluorescence be done simultaneously? Why? Which would you do first? Was homing an efficient strategy for purification of LTR HSC? Is it a generally useful strategy to purify human HSC? How did the authors show that the single injected HSC could self-renew? (serial
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Unformatted text preview: transplantation experiment) How did the authors correlate expression of CD34 and SCA1 with HSC? Do their experiments prove that the bone marrow cells in their pre-purified prep (Fr25, lin-, but-non homed) that express these proteins are HSC? Do the HSC mesodermal derived cells ever give rise to cells of ectodermal origin? of endodermal origin? (at least according to these authors!) How did these authors demonstrate that donor cells were adopting epithelial fates? How are epithelial cells recognized? How are the donor cells recognized? Which epithelial tissue had the most donor cells? What explanation do the authors provide for this Discuss how to culture an ES cell from mice. What about human ES cells? Can adult stem cells be cultured in vitro? (answ- yes, briefly but most are difficult to culture for a long period of time because they spontaneously differentiate)...
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2 - Krause_hili - transplantation experiment How did the...

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