Clonal analysis of single hematopoietic stem cells proliferating under defined conditions in vitro can provide unique insights into their biology and regulation. We have developed a powerful technology for such analyses by establishing a microfluidic system that allows combined automated medium exchange, live-cell immunostaining, and recovery of selected cells. Here we show how this system can be used to investigate the persistence of endothelial protein C receptor (EPCR)-positive cells derived from purified EPCR+ CD150+ CD48- mouse hematopoietic stem cells. These 50% longterm in vivo repopulating cells were stimulated to proliferate under different culture conditions. The results suggest that our microfluidic system can distinguish the clones containing in vivo repopulating progeny of hematopoietic stem cells from the clones lacking such cells. This technology should be useful for the future identification of conditions that optimize hematopoietic stem cell self-renewal divisions, a longstanding challenge for the field.

Clonal analysis of hematopoietic stem cell progeny in microfluidic cell culture arrays

243rd National Meeting (2012)

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