title: Isolation of progenitors that exhibit myogenic/osteogenic bipotency in vitro by fluorescence-activated cell sorting from human fetal muscle.
abstract: Fluorescence-activated cell sorting (FACS) strategies to purify distinct cell types from the pool of fetal human myofiber-associated (hMFA) cells were developed. We demonstrate that cells expressing the satellite cell marker PAX7 are highly enriched within the subset of CD45(-)CD11b(-)GlyA(-)CD31(-)CD34(-)CD56(int)ITGA7(hi) hMFA cells. These CD45(-)CD11b(-)GlyA(-)CD31(-)CD34(-)CD56(int)ITGA7(hi) cells lack adipogenic capacity but exhibit robust, bipotent myogenic and osteogenic activity in vitro and engraft myofibers when transplanted into mouse muscle. In contrast, CD45(-)CD11b(-)GlyA(-)CD31(-)CD34(+) fetal hMFA cells represent stromal constituents of muscle that do not express PAX7, lack myogenic function, and exhibit adipogenic and osteogenic capacity in vitro. Adult muscle likewise contains PAX7(+) CD45(-)CD11b(-)GlyA(-)CD31(-)CD34(-)CD56(int)ITGA7(hi) hMFA cells with in vitro myogenic and osteogenic activity, although these cells are present at lower frequency in comparison to their fetal counterparts. The ability to directly isolate functionally distinct progenitor cells from human muscle will enable novel insights into muscle lineage specification and homeostasis.
authors.affiliation: Howard Hughes Medical Institute, Department of Stem Cell and Regenerative Biology, Harvard University, Harvard Stem Cell Institute, Cambridge, MA 02138, USA ; Joslin Diabetes Center and the Paul F. Glenn Laboratories for the Biological Mechanisms of Aging, Harvard Medical School, Boston, MA 02115, USA ; Vita-Salute San Raffaele University, Milan 20132, Italy.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Castiglioni
authors.affiliation: Howard Hughes Medical Institute, Department of Stem Cell and Regenerative Biology, Harvard University, Harvard Stem Cell Institute, Cambridge, MA 02138, USA ; Joslin Diabetes Center and the Paul F. Glenn Laboratories for the Biological Mechanisms of Aging, Harvard Medical School, Boston, MA 02115, USA ; Department of Pediatric Oncology, Dana Farber Cancer Institute and Division of Pediatric Hematology/Oncology, Children's Hospital, Boston, MA 02115, USA.
authors.forename: Simone
authors.initials: S
authors.lastname: Hettmer
authors.affiliation: Joslin Diabetes Center and the Paul F. Glenn Laboratories for the Biological Mechanisms of Aging, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Matthew D
authors.initials: MD
authors.lastname: Lynes
authors.affiliation: Howard Hughes Medical Institute, Department of Stem Cell and Regenerative Biology, Harvard University, Harvard Stem Cell Institute, Cambridge, MA 02138, USA ; Joslin Diabetes Center and the Paul F. Glenn Laboratories for the Biological Mechanisms of Aging, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Tata Nageswara
authors.initials: TN
authors.lastname: Rao
authors.affiliation: Howard Hughes Medical Institute, Department of Stem Cell and Regenerative Biology, Harvard University, Harvard Stem Cell Institute, Cambridge, MA 02138, USA ; Joslin Diabetes Center and the Paul F. Glenn Laboratories for the Biological Mechanisms of Aging, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Daria
authors.initials: D
authors.lastname: Tchessalova
authors.affiliation: Division of Plastic Surgery, Brigham and Women's Hospital, Boston, MA 02115, USA.
authors.forename: Indranil
authors.initials: I
authors.lastname: Sinha
authors.affiliation: Division of Plastic and Reconstructive Surgery, Department of Surgery, Beth Israel Deaconess Medical Center, Boston, MA 02215, USA.
authors.forename: Bernard T
authors.initials: BT
authors.lastname: Lee
authors.affiliation: Joslin Diabetes Center and the Paul F. Glenn Laboratories for the Biological Mechanisms of Aging, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Yu-Hua
authors.initials: YH
authors.lastname: Tseng
authors.affiliation: Howard Hughes Medical Institute, Department of Stem Cell and Regenerative Biology, Harvard University, Harvard Stem Cell Institute, Cambridge, MA 02138, USA ; Joslin Diabetes Center and the Paul F. Glenn Laboratories for the Biological Mechanisms of Aging, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Amy J
authors.initials: AJ
authors.lastname: Wagers
mesh_terms: D050156:Adipogenesis; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D002454:Cell Differentiation; D019070:Cell Lineage; D002478:Cells, Cultured; D005333:Fetus / Q000166:cytology*; D005434:Flow Cytometry; D006801:Humans; D039001:Integrin alpha Chains / Q000378:metabolism; D051379:Mice; D024510:Muscle Development; D018482:Muscle, Skeletal / Q000166:cytology*; D010012:Osteogenesis; D051763:PAX7 Transcription Factor / Q000378:metabolism; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D014183:Transplantation, Heterologous
chemical_list: D015703:Antigens, CD; D039001:Integrin alpha Chains; D051763:PAX7 Transcription Factor; C497321:PAX7 protein, human; C085725:integrin alpha7