摘要
背景:小鼠骨髓间充质干细胞体外成肌诱导分化效率较低。目的:观察基质金属蛋白酶1在体外对小鼠骨髓间充质干细胞成肌分化的作用。方法:利用差速贴壁法体外分离培养小鼠骨髓间充质干细胞并进行鉴定后,按照不同基质金属蛋白酶1药物处理浓度将小鼠骨髓间充质干细胞分成4组,即10μg/L组、1μg/L组、0.1μg/L组、对照组,给予相应处理后,Real-time定量PCR检测MyoD、Desmin mRNA表达,WesternBlotting检测Desmin蛋白表达。结果与结论:利用差速贴壁法分离培养的小鼠骨髓间充质干细胞形态较一致,表面分子检测示CD29+、Sca-1+、CD45-、CD34-,并具有多向分化潜能;经基质金属蛋白酶1处理后,Real-time定量PCR检测示Desmin、MyoD mRNA表达上调,Western Blotting检测示Desmin蛋白表达上调,并且以上各成肌相关基因、蛋白表达增高程度与基质金属蛋白酶1具有浓度依赖性。提示基质金属蛋白酶1在体外可促进骨髓间充质干细胞向肌肉细胞分化。
BACKGROUND： The myogenic differentiation potential of bone marrow-derived mesenchymal stem cells （MSCs） in vitro from mice is low. OBJECTIVE： To explore the effect of matrix metalloproteinase-1 （MMP-1） on myogenic differentiation of MSCs in vitro from mice. METHODS： After MSCs were isolated and cultured by differential adhesion, as well as identified by cell surface markers and multi-lineage differentiation potential, the cells were assigned to 4 groups according to various MMP-1 concentration gradients （10 IJg/L group, 1 IJg/L group, 0.1 IJg/L group and control group）. The mRNA expressions of MyoD and Desmin were detected by real-time quantitative PCR, and the expression of protein Desmin was tested by Western Blotting. RESULTS AND CONCLUSION： MSCs obtained from differential adhesion were of morphological homogenicity, and they were CD29~, Sca-1 ＋, CD45 and CD34. Additionally, MSCs were capable of multi-lineage differentiation. The mRNA expression of Desmin and MyoD as well as the expression of protein Desmin were up regulated via MMP-1 inducement. Moreover, the up-regulation degree of mRNA and protein expression was concentration-dependent. These suggest that MMP-1 can promote MSCs to differentiate into muscle cells in vitro.
出处
《中国组织工程研究》
CAS
CSCD
2012年第23期4193-4198,共6页
Chinese Journal of Tissue Engineering Research
基金
国家自然科学基金资助项目(30971026
30870852)~~
