胎盘间充质干细胞缺氧培养液对成纤维细胞及巨噬细胞生物学特性的影响

Effect of placenta mesenchymal stem cells hypoxic culture medium on biological characteristics of fibroblasts and macrophages

目的探讨胎盘间充质干细胞(p MSCs)缺氧培养液对成纤维细胞及巨噬细胞的作用及机制。方法提取并鉴定胎盘间充质干细胞,ELISA法检测缺氧时p MSCs分泌胰岛素样生长因子1(IGF-1)水平。台盼蓝染色及划痕实验检测p MSCs条件培养液中成纤维细胞增殖及成纤维细胞、巨噬细胞迁移情况。结果成功提取p MSCs,该细胞表达CD29、CD44,不表达CD31、CD34、CD45及HLA-DR。缺氧p MSCs分泌IGF-1增加。p MSCs缺氧培养液中成纤维细胞增殖快于p MSCs常氧培养液(P<0.05)。加入IGF-1Ab后,细胞增殖减慢(P<0.05)。p MSCs缺氧培养液中成纤维细胞或巨噬细胞迁移快于p MSCs常氧培养液(P<0.05)。加入IGF-1Ab后,细胞迁移减慢(P<0.05)。结论 p MSCs缺氧培养液促进成纤维细胞增殖及迁移,促进巨噬细胞迁移。其可能与p MSCs在缺氧培养液中分泌IGF-1增多相关。

Objective To investigate the effect of placenta mesenchymal stem cells（p MSCs）hypoxic culture medium on fibroblasts and macrophages and discuss its mechanism. Methods p MSCs were extracted and identified. The secretion of insulin-like growth factor-1（IGF-1） by p MSCs in hypoxia was detected by ELISA. The proliferation of fibroblasts and the migration of fibroblasts and macrophages in p MSCs conditioned medium were detected by the method of trypan blue staining and wound scratch healing assay. Results The extracted p MSCs were CD29＋, CD44＋ CD31-,CD34-,CD45-and HLA-DR-.Hypoxia promoted the secretion of IGF-1 by p MSCs. The proliferation of fibroblast in p MSCs hypoxic culture medium（hypo CM） was faster than that in p MSCs normoxic culture medium（nor CM）（P〈0.05）.After the addition of IGF-1 antibody（IGF-1Ab）, proliferation rate was decreased（P 〈0.05）. The migration of fibroblast and macrophage in hypo CM was faster than that in nor CM（P 〈0.05）. After the addition of IGF-1Ab, migration rate was slowed down（P 〈0.05）. Conclusion hypo CM promoted the proliferation and migration of fibroblasts, and promoted the migration of macrophages. Increased IGF-1 in hypo CM was one of the reasons.