HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) All Versions of this Article: 2006-0746v1 25/9/2283    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints/Permissions Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Tamaki, T. Articles by Akatsuka, A. Search for Related Content PubMed PubMed Citation Articles by Tamaki, T. Articles by Akatsuka, A.

TISSUE-SPECIFIC STEM CELLS

Clonal Multipotency of Skeletal Muscle-Derived Stem Cells Between Mesodermal and Ectodermal Lineage

^aMuscle Physiology & Cell Biology Unit,
^bDepartment of Regenerative Medicine, Division of Basic Clinical Science,
^cTeaching & Research Support Center,
^dDepartment of Orthopedics, Division of Surgery, and
^eDepartment of Urology, Division of Surgery, Tokai University School of Medicine, Isehara, Kanagawa, Japan;
^fStem Cell Research, Research Center Japan, Nihon Schering KK, Chuo-ku, Kobe, Japan

Key Words. Tissue-specific stem cells • Myogenic lineage • Neural lineage • Vasculogenic lineage

Correspondence: Tetsuro Tamaki, Ph.D., Muscle Physiology and Cell Biology Unit, Department of Regenerative Medicine, Division of Basic Clinical Science, Tokai University School of Medicine, 143 Shimokasuya, Isehara, Kanagawa 259-1143, Japan. Telephone: +81-463-93-1121 (ext. 2524); Fax: +81-463-95-0961; e-mail: tamaki{at}is.icc.u-tokai.ac.jp

Received November 16, 2006; accepted for publication May 23, 2007.
First published online in STEM CELLS EXPRESS   June 21, 2007.



The differentiation potential of skeletal muscle-derived stem cells (MDSCs) after in vitro culture and in vivo transplantation has been extensively studied. However, the clonal multipotency of MDSCs has yet to be fully determined. Here, we show that single skeletal muscle-derived CD34^–/CD45^– (skeletal muscle-derived double negative [Sk-DN]) cells exhibit clonal multipotency that can give rise to myogenic, vasculogenic, and neural cell lineages after in vivo single cell-derived single sphere implantation and in vitro clonal single cell culture. Muscles from green fluorescent protein (GFP) transgenic mice were enzymatically dissociated and sorted based on CD34 and CD45. Sk-DN cells were clone-sorted into a 96-well plate and were cultured in collagen-based medium with basic fibroblast growth factor and epidermal growth factor for 14 days. Individual colony-forming units (CFUs) were then transplanted directly into severely damaged muscle together with 1 x 10⁵ competitive carrier Sk-DN cells obtained from wild-type mice muscle expanded for 5 days under the same culture conditions using 35-mm culture dishes. Four weeks after transplantation, implanted GFP⁺ cells demonstrated differentiation into endothelial, vascular smooth muscle, skeletal muscle, and neural cell (Schwann cell) lineages. This multipotency was also confirmed by expression of mRNA markers for myogenic (MyoD, myf5), neural (Musashi-1, Nestin, neural cell adhesion molecule-1, peripheral myelin protein-22, Nucleostemin), and vascular (-smooth muscle actin, smoothelin, vascular endothelial-cadherin, tyrosine kinase-endothelial) stem cells by clonal (single-cell derived) single-sphere reverse transcription-polymerase chain reaction. Approximately 70% of clonal CFUs exhibited expression of all three cell lineages. These findings support the notion that Sk-DN cells are a useful tool for damaged muscle-related tissue reconstitution by synchronized vasculogenesis, myogenesis, and neurogenesis.

Disclosure of potential conflicts of interest is found at the end of this article.