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Published online 6 May 2002. doi:10.1083/jcb.200112106
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© The Rockefeller University Press, 0021-9525/2002/5/571 $5.00
The Journal of Cell Biology, Volume 157, Number 4, May 13, 2002 571-577

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Identification of myogenic-endothelial progenitor cells in the interstitial spaces of skeletal muscle



Tetsuro Tamaki1, Akira Akatsuka2, Kiyoshi Ando3,4, Yoshihiko Nakamura3, Hideyuki Matsuzawa3, Tomomitsu Hotta3,4, Roland R Roy5 and V. Reggie Edgerton5,6

1 Department of Physiology, Division of Human Structure and Function
2 Laboratory for Structure and Function Research, Tokai University School of Medicine, Bohseidai, Isehara, Kanagawa 259-1193 Japan
3 Research Center for Genetic Engineeering and Cell Transplantation, Tokai University School of Medicine, Bohseidai, Isehara, Kanagawa 259-1193 Japan
4 Department of Internal Medicine, Tokai University School of Medicine, Bohseidai, Isehara, Kanagawa 259-1193 Japan
5 Brain Research Institute, University of California, Los Angeles, Los Angeles CA 90095
6 Department of Physiological Science, University of California, Los Angeles, Los Angeles CA 90095

Address correspondence to Tetsuro Tamaki, Dept. of Physiology, Division of Human Structure and Function, Tokai University School of Medicine, Bohseidai, Isehara, Kanagawa 259-1193 Japan. Tel.: 81-463-93-1121 x2524. Fax: 81-463-95-0961. E-mail: tamaki{at}is.icc.u-tokai.ac.jp

Putative myogenic and endothelial (myo-endothelial) cell progenitors were identified in the interstitial spaces of murine skeletal muscle by immunohistochemistry and immunoelectron microscopy using CD34 antigen. Enzymatically isolated cells were characterized by fluorescence-activated cell sorting on the basis of cell surface antigen expression, and were sorted as a CD34+ and CD45- fraction. Cells in this fraction were ~94% positive for Sca-1, and mostly negative (<3% positive) for CD14, 31, 49, 144, c-kit, and FLK-1. The CD34+/45- cells formed colonies in clonal cell cultures and colony-forming units displayed the potential to differentiate into adipocytes, endothelial, and myogenic cells. The CD34+/45- cells fully differentiated into vascular endothelial cells and skeletal muscle fibers in vivo after transplantation. Immediately after sorting, CD34+/45- cells expressed only c-met mRNA, and did not express any other myogenic cell-related markers such as MyoD, myf-5, myf-6, myogenin, M-cadherin, Pax-3, and Pax-7. However, after 3 d of culture, these cells expressed mRNA for all myogenic markers. CD34+/45- cells were distinct from satellite cells, as they expressed Bcrp1/ABCG2 gene mRNA (Zhou et al., 2001). These findings suggest that myo-endothelial progenitors reside in the interstitial spaces of mammalian skeletal muscles, and that they can potentially contribute to postnatal skeletal muscle growth.

Key Words: CD34; c-met; Bcrp1; MyoD; Pax-7


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