摘要
背景：前期实验已经证实30%心肌培养上清是诱导大鼠骨髓间充质干细胞分化为心肌样细胞的最佳浓度。目的：在前期实验基础上,观察30%心肌培养上清条件下诱导骨髓间充质干细胞分化为心肌样细胞的时间依赖效应。设计、时间及地点：对比观察,于2007-08/2008-06在徐州市心血管病研究所完成。材料：健康成年SD大鼠,体质量180-220g。方法：分离大鼠骨髓间充质干细胞和心肌细胞并在体外培养纯化。以1×10^8L^-1的细胞密度种植心肌细胞,培养72h后收集上清。将骨髓间充质干细胞的DMEM/F12培养基换为含有30%M199心肌细胞培养上清作为实验组,对照组仍将细胞接种在DMEM/F12培养基上继续培养。主要观察指标：30%心肌细胞培养上清诱导7,14,21d后,应用免疫细胞化学技术检测骨髓间充质干细胞中α-平滑肌肌动蛋白、β-肌动蛋白和心肌特异性肌钙蛋白T的表达。结果：30%心肌细胞培养上清诱导骨髓间充质干细胞后,细胞的形态没有改变,但与对照组相比,α-平滑肌肌动蛋白、β-肌动蛋白和心肌特异性肌钙蛋白T的表达均呈阳性,其中以第14天的蛋白表达量最高（P〈0.01）。结论：30%心肌细胞培养上清诱导骨髓间充质干细胞分化为心肌样细胞的最佳诱导时间是14d。
BACKGROUND： Previous studies have confirmed that 30% myocardial culture supernatant is the optimal concentration of rat bone mesenchymal stem cells （BMSCs） differentiation into cardiomyocyte-like cells （CLCs）. OBJECTIVE： To study the time dependent effects of 30% myocardial culture supernatant on BMSCs differentiated into CLCs. DESIGN, TIME AND SETTING： The controlled experiment was performed at Xuzhou Cardiovascular Disease Institute from August 2007 to June 2008. MATERIALS： Healthy adult Sprague Dawley rats, weighing 180 220 g were selected for this study. METHODS： BMSCs and cardiomyocytes were isolated and cultured in vitro. Cardiomyocytes were cultured at 1×10^8/L for 72 hours. The supernatant liquid was collected and used to induce BMSCs. The DMEM/F12 medium of experimental group was added 30% supernatant liquid of the cardiomyocytes. The control group only used DMEM/F12 to culture BMSCs. MAIN OUTCOME MEASURES： Following 7, 14 and 21 clays of induction in 30% supernatant liquid of the cardiomyocytes, the expression of α - smooth muscle actin, β -actin and Troponin-T was detected by immunocytochemistry. RESULTS： After BMSCs were exposed to 30% supernatant liquid of cardiomyocytes, the morphology of BMSCs did not change. The expression of α -smooth muscle actin, β -actin and Troponin-T were positive compared with the control group, and significantly increased at 14 days （P 〈 0.01 ）. CONCLUSION： The optimal time which BMSCs differentiate into CLCs induced by 30% supernatant liquid of the cardiomyocytes in vitro was 14 days.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2009年第10期1923-1927,共5页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
国家自然科学基金资助项目(30572073)
江苏省自然科学基金项目(BK2005428)
江苏省医学135工程重点人才研究基金(RC2007024)~~
