Ex Vivo Expansion and In Vivo Self-Renewal of Human Muscle Stem Cells.
Adult skeletal muscle stem cells, or satellite cells (SCs), regenerate functional muscle following transplantation into injured or diseased tissue. To gain insight into human SC (huSC) biology, we analyzed transcriptome dynamics by RNA sequencing of prospectively isolated quiescent and activated huSCs. This analysis indicated that huSCs differentiate and lose proliferative potential when maintained in high-mitogen conditions ex vivo. Further analysis of gene expression revealed that p38 MAPK acts in a transcriptional network underlying huSC self-renewal. Activation of p38 signaling correlated with huSC differentiation, while inhibition of p38 reversibly prevented differentiation, enabling expansion of huSCs. When transplanted, expanded huSCs differentiated to generate chimeric muscle and engrafted as SCs in the sublaminar niche with a greater frequency than freshly isolated cells or cells cultured without p38 inhibition. These studies indicate characteristics of the huSC transcriptome that promote expansion ex vivo to allow enhanced functional engraftment of a defined population of self-renewing huSCs.
Paul F. Glenn Laboratories for the Biology of Aging and Department of Neurology and Neurological Sciences, Stanford University School of Medicine, Stanford, CA 94305, USA; Department of Developmental Biology, Stanford University School of Medicine, Stanford, CA 94305, USA.
Gregory W
GW
Charville
Paul F. Glenn Laboratories for the Biology of Aging and Department of Neurology and Neurological Sciences, Stanford University School of Medicine, Stanford, CA 94305, USA; Division of Life Science, The Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong 999077, China.
Tom H
TH
Cheung
Paul F. Glenn Laboratories for the Biology of Aging and Department of Neurology and Neurological Sciences, Stanford University School of Medicine, Stanford, CA 94305, USA.
Bryan
B
Yoo
Paul F. Glenn Laboratories for the Biology of Aging and Department of Neurology and Neurological Sciences, Stanford University School of Medicine, Stanford, CA 94305, USA.
Pauline J
PJ
Santos
Division of Plastic Surgery, Department of Surgery, Stanford University School of Medicine, Stanford, CA 94305, USA.
Gordon K
GK
Lee
Division of Thoracic Surgery, Department of Cardiothoracic Surgery, Stanford University School of Medicine and VA Palo Alto Health Care System, Stanford, CA 94305, USA.
Joseph B
JB
Shrager
Paul F. Glenn Laboratories for the Biology of Aging and Department of Neurology and Neurological Sciences, Stanford University School of Medicine, Stanford, CA 94305, USA; Neurology Service and Rehabilitation Research and Development Center of Excellence, Veterans Affairs Palo Alto Health Care System, Palo Alto, CA 94304, USA. Electronic address: rando@stanford.edu.
Thomas A
TA
Rando
D000328:Adult; D053687:Adult Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D018507:Gene Expression Regulation, Developmental; D053263:Gene Regulatory Networks; D006801:Humans; D051379:Mice; D018482:Muscle, Skeletal / Q000166:cytology* / Q000502:physiology; D012038:Regeneration; D032496:Satellite Cells, Skeletal Muscle / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation; D015398:Signal Transduction; D015533:Transcriptional Activation; D059467:Transcriptome; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
D048051:p38 Mitogen-Activated Protein Kinases