Defining the heterogeneity of skeletal muscle-derived side and main population cells isolated immediately ex vivo.
Defining the heterogeneity of skeletal muscle-derived side and main population cells isolated immediately ex vivo.
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DOI：
10.1002/jcp.21989
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发表时间：
2010-03
影响因子：
5.6
通讯作者：
McLoon, Linda K.
中科院分区：
文献类型：
--
作者：
Kallestad, Kristen M.;McLoon, Linda K.
杜氏肌营养不良症（DMD）的成肌细胞转移治疗在很大程度上由于细胞死亡和移植细胞无法植入患病肌肉而失败。一种尝试富集细胞亚群的方法是Hoechst 33342染料排除测定，产生被认为是祖细胞富集的侧群（SP）和主群（MP）。然而，在体外和移植研究产生了不一致的结果相对于下游后代。骨骼肌来源的MP和SP细胞表达的细胞表面标志物尚未直接离体进行充分表征。使用流式细胞术，MP和SP细胞的特点是基于他们的几个公认的祖细胞抗原的表达。MP和SP群体都是异质的，并且在它们所包含的细胞中重叠。每个群体中细胞的百分比随物种和检查的特定肌肉而变化。MP和SP群体包含卫星和多能祖细胞，基于CD 34、Sca-1、Pax 7和M-钙粘蛋白的表达。因此，使用该程序的分离不能用于预测下游分化结果，并解释了关于这些细胞的相互矛盾的文献。Hoechst染料还导致分选细胞的显著死亡率。由于使用流式细胞术很容易获得确定的亚群，因此基于公认的生肌前体细胞标志物立即离体分选将在移植治疗的细胞同质性方面产生上级结果。
Myoblast transfer therapy for Duchenne muscular dystrophy (DMD) largely fails due to cell death and inability of transplanted cells to engraft in diseased muscles. One method attempting to enrich for cell subpopulations is the Hoechst 33342 dye exclusion assay, yielding a side population (SP) thought to be progenitor enriched and a main population (MP). However, in vitro and transplant studies yielded inconsistent results relative to downstream progeny. Cell surface markers expressed by skeletal muscle-derived MP and SP cells have not been fully characterized directly ex vivo. Using flow cytometry, MP and SP cells were characterized based on their expression of several well-accepted progenitor cell antigens. Both the MP and SP populations are heterogeneous and overlapping in the cells they contain. The percentages of cells in each population vary with species and specific muscle examined. MP and SP populations contain both satellite and multipotent progenitor cells, based on expression of CD34, Sca-1, Pax7, and M-cadherin. Thus, isolation using this procedure cannot be used to predict downstream differentiation outcomes, and explains the conflicting literature on these cells. Hoechst dye also results in significant mortality of sorted cells. As defined subpopulations are easily obtained using flow cytometry, sorting immediately ex vivo based on accepted myogenic precursor cell markers will yield superior results in terms of cell homogeneity for transplantation therapy.
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影响因子：
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