ENZYMATIC LIBERATION OF MYOGENIC CELLS FROM ADULT RAT MUSCLE
ENZYMATIC LIBERATION OF MYOGENIC CELLS FROM ADULT RAT MUSCLE
复制标题
DOI：
10.1002/ar.1091800410
复制
发表时间：
1974-01-01
影响因子：
--
通讯作者：
BISCHOFF, R
中科院分区：
文献类型：
--
作者：
BISCHOFF, R
对胰酶、链霉蛋白酶、胶原酶、木瓜酶和无花果蛋白五种不同的蛋白水解酶从成年大鼠肌肉中释放单核成肌细胞的能力进行了测试。将350~400g大鼠的腿部和大腿肌块与酶孵育1h，用离心法从部分消化的断片中分离出单核细胞。分离的细胞在培养中使用支持胚胎大鼠肌肉分化的程序来测试成肌能力。虽然所有的酶都从肌肉中释放出活细胞，但在体外，只有被胰酶或链霉蛋白酶释放的细胞才能获得肌肉生成。其他三种酶释放成纤维细胞、巨噬细胞和脂肪细胞，但不释放假定的成肌细胞。对消化后的肌肉进行光镜和电子显微镜检查，发现肌源性细胞的酶促释放依赖于去除纤维周围的基底膜。胰酶和链霉蛋白酶能溶解基底膜，而胶原酶、木瓜酶或无花果酶则不能。这些发现表明，在完全成熟的、不能再生的成年肌肉中存在一群肌源性干细胞。这些细胞的频率和释放它们所需的条件表明，它们与肌肉卫星细胞相同。
Five different proteolytic enzymes, trypsin, pronase, collagenase, papain and ficin, were tested for their ability to liberate mononucleated myogenic cells from adult rat muscle. Fragments of leg and thigh muscle from 350–400 gm rats were incubated with enzyme for one hour and mononucleated cells were separated from the partially digested fragments by centrifugation. Myogenic capacity of the isolated cells was tested in culture using procedures known to support differentiation of embryonic rat muscle. Although all the enzymes released viable cells from the muscle, myogenesis in vitro was obtained only from cells liberated by trypsin or pronase. The other three enzymes released fibroblasts, macrophages and fat cells but no presumptive myoblasts. Light and electron microscopic examination of the digested muscle revealed that the enzymatic release of myogenic cells is dependent upon removal of the basement lamina surrounding the fibers. Trypsin and pronase dissolve the basement lamina, whereas collagenase, papain or ficin do not. These findings demonstrate that a population of myogenic stem cells exists in fully mature, non‐regenerating adult muscle. The frequency of these cells and the conditions required for their release suggest that they are identical with the muscle satellite cells.