title: Hematopoietic potential of murine skeletal muscle-derived CD45(-)Sca-1(+)c-kit(-) cells.
abstract: OBJECTIVE
  Somatic stem cells, which are poorly defined in postnatal mammalian tissues, are attractive candidates for examination of stem cell plasticity. Our goal was to determine the identity of neonatal muscle-derived cells that contain hematopoietic potential and to explore the status of CD45 expression on these cells.
  
  
  METHODS
  Skeletal muscle from thighs of 4- to 7-day-old mice was harvested, enzymatically digested, and flow cytometrically sorted to yield CD45(-)Sca-1(+)c-kit(-) cells. These cells were examined in hematopoietic colony-forming assays and competitive repopulation assays, and were expanded ex vivo. Additionally, CD45, c-kit, PU.1, and beta globin major expression was tracked over time in cultured cells to assess the possibility of manipulating stem cell differentiation in vitro.
  
  
  RESULTS
  Freshly isolated CD45(-)Sca-1(+)c-kit(-) cells were devoid of hematopoietic lineage markers and contained no colony-forming activity but displayed superior long-term competitive repopulating ability when compared to freshly isolated muscle-derived CD45(+)Sca-1(+)c-kit(+) cells. CD45(-)Sca-1(+)c-kit(-) cells expanded ex vivo in 5 ng/mL murine stem cell factor, mFlt-3L, and megakaryocyte growth and development factor (MGDF) for 9 days increased their in vivo hematopoietic repopulating potential 5.3-fold relative to fresh cells. Although fresh cells did not transcribe mRNA of several hematopoietic genes, a small fraction of cells cultured for 9 days acquired cell surface c-kit, and only these cells expressed c-kit and PU.1 mRNA and maintained competitive repopulating ability, suggesting at least myeloid and perhaps lymphoid developmental potential.
  
  
  CONCLUSIONS
  Neonatal murine muscle-derived cells expressing the phenotype CD45(-)Sca-1(+) c-kit(-) are putative adult somatic stem cells with in vitro and in vivo hematopoietic differentiation potential.
authors.affiliation: Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis 46202, USA. jchowell@iupui.edu
authors.forename: Jonathan C
authors.initials: JC
authors.lastname: Howell
authors.forename: Mervin C
authors.initials: MC
authors.lastname: Yoder
authors.forename: Edward F
authors.initials: EF
authors.lastname: Srour
mesh_terms: D000818:Animals; D020296:Animals, Congenic; D000831:Animals, Newborn; D000943:Antigens, Differentiation / Q000032:analysis; D000950:Antigens, Ly / Q000032:analysis; D002454:Cell Differentiation; D002469:Cell Separation; D002478:Cells, Cultured / Q000187:drug effects; D003114:Colony-Forming Units Assay; D005434:Flow Cytometry; D005914:Globins / Q000032:analysis; D006412:Hematopoietic Stem Cells / Q000737:chemistry / Q000166:cytology* / Q000187:drug effects; D017493:Leukocyte Common Antigens / Q000032:analysis; D008565:Membrane Proteins / Q000032:analysis / Q000494:pharmacology; D051379:Mice; D008810:Mice, Inbred C57BL; D018482:Muscle, Skeletal / Q000166:cytology*; D011518:Proto-Oncogene Proteins / Q000032:analysis; D019009:Proto-Oncogene Proteins c-kit / Q000032:analysis; D019089:Stem Cell Factor / Q000494:pharmacology; D013926:Thrombopoietin / Q000494:pharmacology; D015534:Trans-Activators / Q000032:analysis; D000099232:Proto-Oncogene Protein Spi-1
chemical_list: D000943:Antigens, Differentiation; D000950:Antigens, Ly; D005914:Globins; D017493:Leukocyte Common Antigens; D008565:Membrane Proteins; D011518:Proto-Oncogene Proteins; D019009:Proto-Oncogene Proteins c-kit; D019089:Stem Cell Factor; D013926:Thrombopoietin; D015534:Trans-Activators; D000099232:Proto-Oncogene Protein Spi-1; C074650:Ly6a protein, mouse; C084881:flt3 ligand protein