DESMIN IS PRESENT IN PROLIFERATING RAT MUSCLE SATELLITE CELLS BUT NOT IN BOVINE MUSCLE SATELLITE CELLS
DESMIN IS PRESENT IN PROLIFERATING RAT MUSCLE SATELLITE CELLS BUT NOT IN BOVINE MUSCLE SATELLITE CELLS
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DOI：
10.1002/jcp.1041490323
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发表时间：
1991-12-01
影响因子：
5.6
通讯作者：
PIERCE, PR
中科院分区：
文献类型：
--
作者：
ALLEN, RE;RANKIN, LL;PIERCE, PR
结蛋白的存在下，其特征在于在培养的大鼠和牛的卫星细胞和其潜在的有用性作为一个标记，用于识别卫星细胞在体外进行了评估。在原代培养中，在大鼠和牛肌管中观察到结蛋白和骨骼肌肌球蛋白的阳性免疫组织化学染色。少量单核细胞（20%的大鼠卫星细胞和5%的牛卫星细胞）肌球蛋白阳性，表明有丝分裂后分化的肌细胞。在牛卫星细胞培养物中，13%的单核细胞结蛋白阳性，而在大鼠卫星细胞培养物中，84%的单核细胞结蛋白阳性。大鼠卫星细胞群培养物和牛卫星细胞克隆密度培养物用H-3-胸苷脉冲，放射自显影数据显示> 94%的分裂大鼠细胞为结蛋白阳性，表明结蛋白在增殖的大鼠卫星细胞中合成。然而，没有结蛋白被认为是在细胞中标记的胸苷在牛卫星细胞克隆。克隆密度培养物的分析表明，只有14%的单核细胞在牛卫星细胞集落的结蛋白阳性，而98%的细胞在大鼠卫星细胞集落的结蛋白阳性。两个物种的成纤维细胞集落均为结蛋白阴性。为了进一步检查卫星细胞分化和结蛋白表达之间的关系，在铺板时向培养基中加入5-溴-2 '-脱氧尿苷（BrdU）以抑制分化。在大鼠和牛培养物中，融合受到抑制，细胞继续分裂。在牛培养物中发现极少数结蛋白阳性细胞，但在大鼠培养物中超过90%的细胞对结蛋白染色呈阳性。结蛋白和肌节肌球蛋白的存在下，也进行了评价再生大鼠胫骨前布比卡因注射后5天。在再生区的肌肉许多结蛋白阳性细胞，只有少数细胞染色阳性骨骼肌肌球蛋白。应用结蛋白染色大鼠卫星细胞生长试验表明，大鼠卫星细胞培养在含血清的培养基中污染成纤维细胞的水平，范围从约5%，在24小时的培养物中的成熟培养物中的15%。在确定成分培养基中，4天培养物含有约95%至98%的结蛋白阳性卫星细胞。通过结蛋白染色评估胰岛素样生长因子I（IGF-I）、碱性成纤维细胞生长因子（bFGF）和转化生长因子β（TGF-β）组合对大鼠卫星细胞增殖和分化的影响，发现结果与先前使用常规细胞染色和计数技术获得的结果一致（艾伦and Boxhorn，1989）。我们的实验表明，在卫星细胞中的结蛋白表达的模式不同，大鼠和牛，结蛋白可以是一个有用的标记培养的大鼠卫星细胞。
The presence of desmin was characterized in cultured rat and bovine satellite cells and its potential usefulness as a marker for identifying satellite cells in vitro was evaluated. In primary cultures, positive immunohistochemical staining for desmin and skeletal muscle myosin was observed in rat and bovine myotubes. A small number of mononucleated cells (20% of rat satellite cells and 5% of bovine satellite cells) were myosin-positive, indicative of post-mitotic differentiated myocytes. In bovine satellite cell cultures 13% of the mononucleated cells were desmin-positive, while 84% of the mononucleated cells in rat satellite cell cultures were desmin-positive. Rat satellite cell mass cultures and bovine satellite cell clonal density cultures were pulsed with H-3-thymidine, and autoradiographic data revealed that > 94% of dividing rat cells were desmin-positive, suggesting that desmin is synthesized in proliferating rat satellite cells. However, no desmin was seen in cells that incorporated labeled thymidine in bovine satellite cell clones. Analysis of clonal density cultures revealed that only 14% of the mononucleated cells in bovine satellite cell colonies were desmin-positive, whereas 98% of the cells in rat satellite cell colonies were desmin-positive. Fibroblast colonies from both species were desmin-negative. In order to further examine the relationship between satellite cell differentiation and desmin expression, 5-bromo-2'-deoxyuridine (BrdU) was added to culture medium at the time of plating to inhibit differentiation. Fusion was inhibited in rat and bovine cultures, and cells continued to divide. Very few desmin-positive cells were found in bovine cultures, but greater than 90% of the cells in rat cultures stained positive for desmin. The presence of desmin and sarcomeric myosin was also evaluated in regenerating rat tibialis anterior five days after bupivacaine injection. In regenerating areas of the muscle many desmin-positive cells were present, and only a few cells stained positive for skeletal muscle myosin. Application of desmin staining to rat satellite cell growth assays indicated that rat satellite cells cultured in serum-containing medium were contaminated with fibroblasts at levels that ranged from approximately 5% in 24 hr cultures to 15% in mature cultures. In defined medium 4 day cultures contain approximately 95% to 98% desmin-positive satellite cells. The effects of combinations of insulin-like growth factor I (IGF-I), basic fibroblast growth factor (bFGF), and transforming growth factor beta (TGF-beta) on rat satellite cell proliferation and differentiation were assessed by desmin staining, and results were found to be consistent with results obtained previously using conventional cell staining and counting techniques (Allen and Boxhorn, 1989). Our experiments indicate that the pattern of desmin expression in satellite cells differs between rat and bovine and that desmin can be a useful marker for cultured rat satellite cells.