Sorting live stem cells based on Sox2 mRNA expression.

Hans M Larsson, Seung Tae Lee, Marta Roccio, Diana Velluto, Matthias P Lutolf, Peter Frey, Jeffrey A Hubbell

PloS one 2012

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While cell sorting usually relies on cell-surface protein markers, molecular beacons (MBs) offer the potential to sort cells based on the presence of any expressed mRNA and in principle could be extremely useful to sort rare cell populations from primary isolates. We show here how stem cells can be purified from mixed cell populations by sorting based on MBs. Specifically, we designed molecular beacons targeting Sox2, a well-known stem cell marker for murine embryonic (mES) and neural stem cells (NSC). One of our designed molecular beacons displayed an increase in fluorescence compared to a nonspecific molecular beacon both in vitro and in vivo when tested in mES and NSCs. We sorted Sox2-MB(+)SSEA1(+) cells from a mixed population of 4-day retinoic acid-treated mES cells and effectively isolated live undifferentiated stem cells. Additionally, Sox2-MB(+) cells isolated from primary mouse brains were sorted and generated neurospheres with higher efficiency than Sox2-MB(-) cells. These results demonstrate the utility of MBs for stem cell sorting in an mRNA-specific manner.

Citation

Hans M Larsson, Seung Tae Lee, Marta Roccio, Diana Velluto, Matthias P Lutolf, Peter Frey, Jeffrey A Hubbell. Sorting live stem cells based on Sox2 mRNA expression. PloS one. 2012;7(11):e49874