title: Synchronized reconstitution of muscle fibers, peripheral nerves and blood vessels by murine skeletal muscle-derived CD34(-)/45 (-) cells.
abstract: In order to establish the practical isolation and usage of skeletal muscle-derived stem cells (MDSCs), we determined reconstitution capacity of CD34(-)/CD45(-) (Sk-DN) cells as a candidate somatic stem cell source for transplantation. Sk-DN cells were enzymatically isolated from GFP transgenic mice (C57/BL6N) skeletal muscle and sorted using fluorescence activated cell sorting (FACS), and expanded by collagen gel-based cell culture with bFGF and EGF. The number of Sk-DN cells was small after sorting (2-8 x 10(4)); however, the number increased 10-20 fold (2-16 x 10(5)) after 6 days of expansion culture, and the cells maintained immature state and multipotency, expressing mRNAs for mesodermal and ectodermal cell lineages. Transplantation of expanded Sk-DN cells into the severe muscle damage model (C57/BL6N wild-type) resulted in the synchronized reconstitution of blood vessels, peripheral nerves and muscle fibers following significant recovery of total muscle mass (57%) and contractile function (55%), whereas the non-cell-transplanted control group showed around 20% recovery in both factors. These reconstitution capacities were supported by the intrinsic plasticity of Sk-DN cells that can differentiate into muscular (skeletal muscle), vascular (pericyte, endothelial cell and smooth muscle) and peripheral nerve (Schwann cells and perineurium) cell lineages that was revealed by transplantation to non-muscle tissue (beneath renal capsule) and fluorescence in situ hybridization (FISH) analysis.
authors.affiliation: Muscle Physiology and Cell Biology Unit, Tokai University School of Medicine, Shimokasuya-143, Isehara, Kanagawa, 259-1143, Japan. tamaki@is.icc.u-tokai.ac.jp
authors.forename: Tetsuro
authors.initials: T
authors.lastname: Tamaki
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Okada
authors.forename: Yoshiyasu
authors.initials: Y
authors.lastname: Uchiyama
authors.forename: Kayoko
authors.initials: K
authors.lastname: Tono
authors.forename: Maki
authors.initials: M
authors.lastname: Masuda
authors.forename: Mika
authors.initials: M
authors.lastname: Wada
authors.forename: Akio
authors.initials: A
authors.lastname: Hoshi
authors.forename: Akira
authors.initials: A
authors.lastname: Akatsuka
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000235:genetics / Q000378:metabolism*; D002454:Cell Differentiation; D002478:Cells, Cultured; D005260:Female; D005786:Gene Expression Regulation; D017404:In Situ Hybridization, Fluorescence; D017493:Leukocyte Common Antigens / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D016253:Microscopy, Immunoelectron; D024510:Muscle Development; D018485:Muscle Fibers, Skeletal / Q000378:metabolism* / Q000648:ultrastructure; D018482:Muscle, Skeletal / Q000098:blood supply* / Q000378:metabolism*; D010525:Peripheral Nerves / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats
chemical_list: D018952:Antigens, CD34; D012333:RNA, Messenger; D017493:Leukocyte Common Antigens