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First published online 6 October 2004
doi: 10.1242/dev.01395
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1 Tissue Stem Cell Research Team, Mitsubishi Kagaku Institute of Life Sciences,
11 Minamiooya, Machida, Tokyo 194-8511, Japan
2 Fine Structure Analysis Section, Mitsubishi Kagaku Institute of Life Sciences,
11 Minamiooya, Machida, Tokyo 194-8511, Japan
3 Kitasato University, School of Science, Department of Biosciences, 1-15-1
Kitasato, Sagamihara, Kanagawa 228-8555, Japan
* Author for correspondence (e-mail: nao{at}libra.ls.m-kagaku.co.jp)
Accepted 11 August 2004
Recent studies have demonstrated that a distinct subpopulation with stem cell-like characteristics in myoblast culture is responsible for new muscle fiber formation after intramuscular transplantation. The identification and isolation of stem-like cells would have significant implications for successful myogenic cell transfer therapy in human muscle disorders. Using a clonal culture system for mouse muscle satellite cells, we have identified two cell types, designated `round cells' and `thick cells', in clones derived from single muscle satellite cells that have been taken from either slow or fast muscle. Clonal analysis of satellite cells revealed that the round cells are immediate descendants of quiescent satellite cells in adult muscle. In single-myofiber culture, round cells first formed colonies and then generated progeny, thick cells, that underwent both myogenic and osteogenic terminal differentiation under the appropriate culture conditions. Thick cells, but not round cells, responded to terminal differentiation-inducing signals. Round cells express Pax7, a specific marker of satellite cells, at high levels. Myogenic cell transfer experiments showed that round cells reconstitute myofibers more efficiently than thick cells. Furthermore, round cells restored dystrophin in myofibers of mdx nude mice, even when as few as 5000 cells were transferred into the gastrocnemius muscle. These results suggest that round cells are satellite-cell descendants with stem cell-like characteristics and represent a useful source of donor cells to improve muscle regeneration.
Key words: Myoblast transfer therapy, Muscle regeneration, Muscle satellite cell
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