Growth factors improve the in vivo migration of human skeletal myoblasts by modulating their endogenous proteolytic activity
Growth factors improve the in vivo migration of human skeletal myoblasts by modulating their endogenous proteolytic activity
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DOI：
10.1097/01.tp.0000131175.60047.eb
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发表时间：
2004-06-15
期刊：
影响因子：
6.2
通讯作者：
El Fahime, E
中科院分区：
文献类型：
--
作者：
Lafreniere, JF;Mills, P;El Fahime, E
背景与成肌细胞移植临床应用相关的一个主要技术问题是移植细胞的迁移性差。在这项研究中，我们研究了一种新的生理方法，包括共注射运动因子胰岛素生长因子（IGF）-1或碱性成纤维细胞生长因子（bFGF），以提高人骨骼肌成肌细胞的迁移。在这些因素可以诱导细胞迁移过程的不同方式中，我们研究了它们增强细胞内源性蛋白水解活性的能力，这将有助于移植细胞通过细胞外基质迁移。在体外，成肌细胞与bFGF或IGF-1共孵育。生长因子对细胞迁移的影响进行了评估，使用入侵室，并通过酶谱法，蛋白质印迹法，和逆转录聚合酶链反应，其对蛋白水解系统的影响进行了评估。在体内，成肌细胞与生长因子共注射，并使用微管技术评估肌内迁移能力。在体外，IGF-1或bFGF的存在下显着增强明胶酶基质金属蛋白酶-9的表达，并集中在细胞膜上的纤溶系统的活动。在体外和体内，bFGF和IGF-1都表现出强大的趋化动力学潜力，并改善人成肌细胞的迁移。此外，使用特异性抑制剂（BB 94或阿米洛利）证实了某些蛋白酶在体内增强迁移中的意义。这些结果表明，IGF-1或bFGF与人成肌细胞共注射增加了它们的蛋白水解活性，从而增加了它们的迁移能力。这项研究可能有助于开发方法，将减少注射部位的数量，用于治疗杜氏肌营养不良症患者。
Background. A main technological problem related to the clinical application of myoblast transplantation is the poor migration of transplanted cells. In this study, we investigated a new physiologic approach that consists of coinjecting motogenic factors insulin growth factor (IGF)-1 or basic fibroblast growth factor (bFGF) to enhance the migration of human skeletal myoblasts. Among the different ways by which those factors can induce the cell migration processes, we investigated their capacity to enhance cell endogenous proteolytic activity that will help transplanted cells to migrate through the extracellular matrix.Methods. In vitro, myoblasts were coincubated with bFGF or IGF-1. Growth factors effects on cell migration were evaluated using invasion chambers, and their effects on proteolytic systems were evaluated by zymography, Western blot, and reverse transcription polymerase chain reaction. In vivo, myoblasts were coinjected with growth factors and the intramuscular migration capacity was assessed using the microtube technique.Results. In vitro, the presence of IGF-1 or bFGF significantly enhanced the expression of the gelatinase matrix metalloproteinase-9 and focalized the fibrinolytic system activity at the cell membrane. In vitro and in vivo, both bFGF and IGF-1 showed strong chemokinetic potentials and improved the migration of human myoblasts. Moreover, the implication some proteinases in the in vivo enhanced migration was confirmed using specific inhibitors (BB94 or amiloride).Conclusions. These results suggest that IGF-1 or bFGF coinjection with human myoblasts increased their proteolytic activities and consequently their migratory capacity. This study may help to develop approaches that will reduce the number of injection sites for the treatment of Duchenne muscular dystrophy patients.