title: A novel approach to collecting satellite cells from adult skeletal muscles on the basis of their stress tolerance.
abstract: Stem cells are generally collected using flow cytometry, but this method is not applicable when the cell surface marker is not well determined. Satellite cells, which are skeletal muscle stem cells, have the ability to regenerate damaged muscles and are expected to be applicable for treatment of muscle degeneration. Although the transcription factor Pax7 is a known specific marker of satellite cells, it is not located on the cell surface and therefore flow cytometry is not directly applicable. In the present study, we turned our attention to the stress tolerance of adult stem cells, and we propose long-term trypsin incubation (LTT) as a novel approach to collecting satellite cells from mouse and human skeletal muscles. LTT led to a remarkable increase in the ratio of Pax7(+) cells that retain normal myogenic stem cell function. In particular, human Pax7(+) cells made up approximately 30% of primary cultured cells, whereas after LTT, the ratio of Pax7(+) cells increased up to ∼80%, and the ratio of Pax7(+) and/or MyoD(+) myogenic cells increased to ∼95%. Once transplanted, LTT-treated cells contributed to subsequent muscle regeneration following repetitive muscle damage without additional cell transplantation. The stress tolerance of Pax7(+) cells is related to heat shock protein 27 and αB-crystallin, members of the small heat shock protein family. This approach, based on the stress resistance of adult stem cells, is a safe and inexpensive method of efficiently collecting human satellite cells and may also be used for collecting other tissue stem cells whose surface marker is unknown.
authors.affiliation: Department of Stem Cell Biology and Histology, Tohoku University Graduate School of Medicine, Sendai, Miyagi, Japan. tshigemoto@med.tohoku.ac.jp
authors.forename: Taeko
authors.initials: T
authors.lastname: Shigemoto
authors.forename: Yasumasa
authors.initials: Y
authors.lastname: Kuroda
authors.forename: Shohei
authors.initials: S
authors.lastname: Wakao
authors.forename: Mari
authors.initials: M
authors.lastname: Dezawa
mesh_terms: D000369:Aged, 80 and over; D000818:Animals; D002469:Cell Separation / Q000379:methods*; D005260:Female; D055551:HSP27 Heat-Shock Proteins / Q000378:metabolism; D006360:Heat-Shock Proteins; D006801:Humans; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018832:Molecular Chaperones; D032342:Muscle Cells / Q000166:cytology* / Q000378:metabolism; D018482:Muscle, Skeletal / Q000166:cytology; D017570:MyoD Protein / Q000378:metabolism; D051763:PAX7 Transcription Factor / Q000378:metabolism; D012038:Regeneration / Q000502:physiology; D032496:Satellite Cells, Skeletal Muscle / Q000166:cytology* / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D013312:Stress, Physiological / Q000502:physiology*; D014357:Trypsin / Q000494:pharmacology; D038203:alpha-Crystallin B Chain / Q000378:metabolism
chemical_list: D055551:HSP27 Heat-Shock Proteins; C057725:HSPB1 protein, human; D006360:Heat-Shock Proteins; C486699:Hspb2 protein, mouse; D018832:Molecular Chaperones; D017570:MyoD Protein; C057363:MyoD1 myogenic differentiation protein; D051763:PAX7 Transcription Factor; C497321:PAX7 protein, human; C497322:Pax7 protein, mouse; D038203:alpha-Crystallin B Chain; D014357:Trypsin
keywords: Adult stem cells; Muscle stem cells; Somatic cell therapy; Tissue-specific stem cells