title: Sorting live stem cells based on Sox2 mRNA expression.
abstract: 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.
authors.affiliation: Laboratory for Regenerative Medicine and Pharmacobiology, Institute for Bioengineering, School of Life Sciences and School of Engineering, Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.
authors.forename: Hans M
authors.initials: HM
authors.lastname: Larsson
authors.forename: Seung Tae
authors.initials: ST
authors.lastname: Lee
authors.forename: Marta
authors.initials: M
authors.lastname: Roccio
authors.forename: Diana
authors.initials: D
authors.lastname: Velluto
authors.forename: Matthias P
authors.initials: MP
authors.lastname: Lutolf
authors.forename: Peter
authors.initials: P
authors.lastname: Frey
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Hubbell
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002469:Cell Separation / Q000379:methods; D005434:Flow Cytometry; D015870:Gene Expression; D051379:Mice; D058953:Neural Stem Cells / Q000166:cytology / Q000378:metabolism; D055748:SOXB1 Transcription Factors / Q000235:genetics* / Q000378:metabolism*; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism*
chemical_list: D055748:SOXB1 Transcription Factors; C527783:Sox2 protein, mouse