Increased myogenic potential and fusion of matrilysin-expressing myoblasts transplanted in mice
Increased myogenic potential and fusion of matrilysin-expressing myoblasts transplanted in mice
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DOI：
10.1177/096368979900800502
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发表时间：
1999-09-01
影响因子：
3.3
通讯作者：
Tremblay, JP
中科院分区：
文献类型：
--
作者：
Caron, NJ;Asselin, I;Tremblay, JP
成肌细胞移植在临床试验中的成功部分受到移植细胞在注射部位外分散程度低的限制。我们的研究小组之前报道过，用豆蛋白A（一种金属蛋白酶产生的刺激剂）培养成肌细胞，增加了它们的迁移。一些证据也表明肌肉细胞融合涉及金属蛋白酶敏感机制。为了确定金属蛋白酶表达的增加是否对移植后成肌细胞的分裂和分散有影响，我们产生了表达人基质溶素（MMP-7）的成肌细胞系。将基质溶素表达载体稳定转染到TnILacZ永不朽小鼠成肌细胞克隆中，获得表达mmp -7的成肌细胞。表达母溶素的成肌细胞的体外融合指数显著提高，形成的肌管比对照细胞系多7倍（p < 0.001），比成不朽细胞亲本克隆多3倍（p < 0.001）。单位点移植表达基质溶素的成肌细胞比对照细胞系产生更多的纤维（p < 0.001），在更大的表面（p < 0.001）。将表达基质溶素的成肌细胞与正常人成肌细胞共移植到SCID小鼠体内，人肌营养不良蛋白阳性纤维和肌管的数量增加了6倍。虽然没有观察到成肌细胞在注射部位外的迁移明显增加，但我们的研究结果表明，金属蛋白酶活性可以提高体外成肌细胞的成肌潜能和体内成肌细胞与宿主纤维的融合。MMP-7的表达可能有助于提高成肌细胞移植的成功率。
The success of myoblast transplantation in clinical trials has been limited in part by the low dispersion of grafted cells outside the injection site. Our research group previously reported that the culture of myoblasts with concanavalin A, a stimulator of metalloproteinase production, increased their migration. Several lines of evidence also suggested that muscle cell fusion involves metalloproteinase-sensitive mechanisms. To determine whether the increased expression of metalloproteinases had an influence on myoblast fission and dispersion through the muscle following transplantation, we generated a myoblast cell line expressing human matrilysin (MMP-7). The MMP-7-expressing myoblasts were obtained by the stable transfection of a matrilysin expression vector in a TnILacZ immortomouse myoblast clone. Matrilysin-expressing myoblasts showed a highly increased in vitro fusion index, forming seven times (p < 0.001) more myotubes than the control cell line and three times (p,< 0.001) more myotubes than the Immortomyoblast parental clone. Single-site transplantation of matrilysin-expressing myoblasts generated more fibers (p < 0.001), over a greater surface (p < 0.001) than the control cell line. The cotransplantation of matrilysin-expressing myoblasts and of normal human myoblasts in SCID mice increased the number of human dystrophin-positive fibers and myotubes by sixfold. Although no significant increased migration of myoblasts outside the injection sites was observed, our results show that the metalloproteinase activity can improve the myogenic potential of myoblasts in vitro and the fusion of myoblasts with host fibers in vivo. MMP-7 expression may be useful in increasing myoblast transplantation success.