人胎盘间充质干细胞微环境对心肌细胞增殖的影响  

Effects of microenvironment produced by human pMSC on the proliferation of cardiomyocytes

作　　者：章毅 陈亮 李萍 许惠利 王哲 伍婷 张澄宇 ZHANG Yi CHEN Liang LI Ping XU Hui-li WANG Zhe WU Ting ZHANG Cheng-yu(Shanghai Cord Blood Bank/Shanghai Stem Cell Technology Co., Ltd./China Stem Cell Group Co., Ltd., Shanghai 200051, China)

机构地区：[1]上海市脐带血造血干细胞库/上海市干细胞技术有限公司/中国干细胞集团有限公司,200051

出　　处：《中国医药生物技术》2017年第2期149-156,共8页Chinese Medicinal Biotechnology

基　　金：上海市自然科学基金(16ZR1429200);上海市科委研发平台专项(16DZ2293000);嘉定区工程技术研究中心项目;2016年上海市专利试点企业项目

摘　　要：目的观察胎盘间充质干细胞微环境对心肌细胞增殖的影响。方法用胎盘间充质干细胞培养基上清模拟胎盘间充质干细胞微环境,与H9C2心肌细胞transwell共培养(微环境组),观察其对H9C2细胞的生长状态、细胞增殖影响情况。同时设置10%FBS的DMEM为空白对照、pMSC/H9C2transwell共培养为阳性对照(pMSC组),倒置显微镜观察心肌细胞增殖状态;共培养72 h后将H9C2胰酶消化按5×10~6/ml每孔进行96孔板铺板,分别在24和48 h进行CCK8细胞增殖检测。结果与对照组相比,胎盘间充质干细胞培养基上清与心肌共培养72 h后心肌细胞密度达到80%以上,显著高于对照组,与pMSC组无明显差异。共培养处理后的心肌细胞培养24和48 h增殖结果均显示,微环境组与pMSC组心肌细胞增殖高于对照组,差异极显著(P<0.01),24 h增殖结果显示pMSC组与微环境组心肌细胞增殖差异极显著(P<0.01),而48 h增殖结果显示pMSC组与微环境组心肌细胞增殖差异不显著(P>0.05)。结论胎盘间充质干细胞微环境与胎盘间充质干细胞对心肌细胞的增殖均具有显著的促进作用,提示胎盘间充质干细胞微环境有望部分替代胎盘间充质干细胞的心肌损伤修复治疗功能。Objective To investigate the effect of microenvironment created by human placental mesenchymal stem cells （pMSC） on the proliferation of cardiomyocytes.Methods The cultured medium of pMSC was used to simulate the microenvironment produced by pMSC and co-culture with cardiomyocytes （H9C2） through the method of transwell experiment （the microenvironment group）. At the same time, the DMEM medium containing 10% FBS and pMSC/H9C2 transwell group were treated as blank and positive control, respectively. After 72 h, the cell proliferation and morphology were observed under microscope. Then H9C2 cells were digested by trypsin to plant 96-well plate with cell concentration of 5 ＆#215; 106/ml. CCK8 test was carried out at 24 and 48 h after cell seeding, respectively.Results Compared with the control group, H9C2 cells grew more rapidly in the microenvironment group after 72 h of co-culture. The density of H9C2 reached 80%, significantly higher than that in the control group. There was no significant difference between the medium group and pMSC group. CCK8 cell proliferation assay at 24 and 48 h showed that the values of the microenvironment group and the pMSC group were significant higher than that in the control （P 〈 0.01）. Significant difference was observed between the microenvironment group and pMSC group at 24 h （P〈 0.01）, while no difference observed between the two groups at 48 h （P〉 0.05）.Conclusion Both microenvironment and pMSC can promote the cell proliferation of H9C2. The results imply that the microenvironment of pMSC is expected to partly take place of the repair function of pMSC in the myocardial injury field.

关 键 词：肌细胞 心脏 细胞微环境 胎盘间充质干细胞 

分 类 号：R54[医药卫生—心血管疾病]