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      Expression of Cd34 and Myf5 Defines the Majority of Quiescent Adult Skeletal Muscle Satellite Cells


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          Skeletal muscle is one of a several adult post-mitotic tissues that retain the capacity to regenerate. This relies on a population of quiescent precursors, termed satellite cells. Here we describe two novel markers of quiescent satellite cells: CD34, an established marker of hematopoietic stem cells, and Myf5, the earliest marker of myogenic commitment. CD34 +ve myoblasts can be detected in proliferating C2C12 cultures. In differentiating cultures, CD34 +ve cells do not fuse into myotubes, nor express MyoD. Using isolated myofibers as a model of synchronous precursor cell activation, we show that quiescent satellite cells express CD34. An early feature of their activation is alternate splicing followed by complete transcriptional shutdown of CD34. This data implicates CD34 in the maintenance of satellite cell quiescence.

          In heterozygous Myf5 nlacZ/ + mice, all CD34 +ve satellite cells also express β-galactosidase, a marker of activation of Myf5, showing that quiescent satellite cells are committed to myogenesis. All such cells are positive for the accepted satellite cell marker, M-cadherin. We also show that satellite cells can be identified on isolated myofibers of the myosin light chain 3F- nlacZ-2E mouse as those that do not express the transgene. The numbers of satellite cells detected in this way are significantly greater than those identified by the other three markers.

          We conclude that the expression of CD34, Myf5, and M-cadherin defines quiescent, committed precursors and speculate that the CD34 −ve, Myf5 −ve minority may be involved in maintaining the lineage-committed majority.

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          Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.

          A new method of total RNA isolation by a single extraction with an acid guanidinium thiocyanate-phenol-chloroform mixture is described. The method provides a pure preparation of undegraded RNA in high yield and can be completed within 4 h. It is particularly useful for processing large numbers of samples and for isolation of RNA from minute quantities of cells or tissue samples.
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            Liver regeneration after the loss of hepatic tissue is a fundamental parameter of liver response to injury. Recognized as a phenomenon from mythological times, it is now defined as an orchestrated response induced by specific external stimuli and involving sequential changes in gene expression, growth factor production, and morphologic structure. Many growth factors and cytokines, most notably hepatocyte growth factor, epidermal growth factor, transforming growth factor-alpha, interleukin-6, tumor necrosis factor-alpha, insulin, and norepinephrine, appear to play important roles in this process. This review attempts to integrate the findings of the last three decades and looks toward clues as to the nature of the causes that trigger this fascinating organ and cellular response.
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              Stem cells: units of development, units of regeneration, and units in evolution.


                Author and article information

                J Cell Biol
                The Journal of Cell Biology
                The Rockefeller University Press
                11 December 2000
                : 151
                : 6
                : 1221-1234
                [a ]Muscle Cell Biology Group, Medical Research Council Clinical Sciences Centre, Imperial College School of Medicine, Hammersmith Hospital, London, W12 ONN United Kingdom
                [b ]Centre National de la Recherche Scientifique Unité de Recherche Associée 1947, Département de Biologie Moléculaire, Institut Pasteur, 75724 Paris Cedex 15, France
                [c ]Physiologisches Institut II der Universitat Bonn, Neurophysiologie, D-53111, Bonn, Germany
                © 2000 The Rockefeller University Press
                : 10 April 2000
                : 11 September 2000
                : 10 October 2000
                Original Article

                Cell biology
                satellite cell,myod,myf5,skeletal muscle,cd34
                Cell biology
                satellite cell, myod, myf5, skeletal muscle, cd34


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