干细胞因子促进骨髓间充质干细胞向心肌细胞的分化(英文)  

Differentiation of mesenchymal stem cells into myocardial cells promoted by stem cell factors

作　　者：鲍翠玉[1,2] 郭军[1] 马业新[3] 郑敏[1] 

机构地区：[1]咸宁学院心血管疾病研究所 [2]华中科技大学同济医学院附属同济医院心内科,湖北省武汉市430030 [3]华中科技大学同济医学院附属同济医院心内科

出　　处：《中国临床康复》2006年第41期180-182,I0006,共4页Chinese Journal of Clinical Rehabilitation

基　　金：湖北省教育厅科研计划项目(2004Q001);湖北省自然科学基金(2005ABA085)~~

摘　　要：背景:骨髓间质干细胞体内、外均能分化为心肌细胞,但数量少且定向分化率较低。目的:探讨干细胞因子促进骨髓间充质干细胞向心肌细胞分化的作用。设计:开放性实验。单位:咸宁学院心血管疾病研究所,华中科技大学同济医学院附属同济医院心内科。材料:实验于2003-10/2004-08在咸宁学院心血管疾病研究所实验中心完成。选取出生1～2d的SD新生大鼠20只,用于心肌细胞的培养。另选取清洁级成年SD大鼠25只,随机数字表法分为干细胞因子组、空白对照组,12只/组,剩余1只大鼠用于骨髓间充质干细胞的提取、培养及纯化。方法:①在共培养前用4,6-联脒-2-苯基吲哚标记骨髓间充质干细胞。干细胞因子组对大鼠进行体内动员,连续5d皮下注射重组鼠干细胞因子,20μg/(kg·d),然后提取其骨髓间充质干细胞于心肌细胞培养第3天进行共培养。空白对照组皮下注射相等剂量的生理盐水,未施加任何干预的骨髓间充质干细胞于心肌细胞培养第3天进行共培养。②用数码显微摄像及免疫荧光技术分别记录和检测MHCα/β、肌钙蛋白T的表达,测定4,6-联脒-2-苯基吲哚-骨髓间充质干细胞向心肌细胞分化的百分率。主要观察指标:①骨髓间充质干细胞生长情况。②检测4,6-联脒-2-苯基吲哚对骨髓间充质干细胞的核标记率。③拟做共培养的心肌细胞活力、纯度分析。④共培养后骨髓间充质干细胞向心肌细胞的分化情况。结果:①骨髓细胞悬液接种于塑料培养皿,细胞呈圆形散在分布于瓶底,24h换液后可见稀少的长梭形贴壁细胞,4d后梭形贴壁细胞开始增多,进入对数增长期,8～12d达到80%的融合。传代细胞增殖更加迅速,随着换液与传代,骨髓间充质干细胞逐渐得到纯化。②4,6-联脒-2-苯基吲哚对骨髓间充质干细胞的胞核标记率为100%。③心肌细胞体外培养第3天,搏动细胞的百分比为63%,搏动频率4BACKGROUND： Mesenchymal stem cells （MSCs） can differentiate into myocardial cells in vitro and in vivo, but the amount is small and directed differentiation rate is low. OBJECTIVE： To explore the effect of stem cell factor （SCF） on promotion of MSCs differentiating into myocardial cells. DESIGN： Opening experiment. SETTING： Institute of Cardiovascular Disease, Xianning College and Department of Cardiology, Tongji Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology. MATERIALS： The experiment was performed at the Experimental Center of Institute of Cardiovascular Disease, Xianning College from October 2003 to August 2004. A total of 20 infant SD rats aged 1-2 days were selected for culture of myocardial cells. Another 25 clean adult SD rats were selected and randomly divided into SCF group, blank control group with 12 rats in each group. The left one rat was used for MSCs extraction, culture and purification. METHODS： （1）MSCs were labeled with 4＇, 6-diamidino-2-phenylindole, dihydrochloride （DAPI） before co-cuhure. SCF was given into rats of the SCF group by subcutaneous injection successively for 5 days, 20 μg/kg per day. Isolated MSCs were co-cultured with myocardial cells that had been cultured for 3 days. Saline of the same volume was given in the blank control group by subcutaneous injection. MSCs without any intervention were co-cultured with myocardial cells that had been cultured for 3 days. （2）Expressions of MHC α/β and troponin T were recorded and measured with digital micro-camera shot and immunofluorescence technique, respectively. Percentage of DAPI labeled MSCs differentiating into myocardial cells was measured. MAIN OUTCOME MEASURES： （1）Growth of MSCs, （2）detection of labeling rate of DAPI on MSCs, （3）analysis of activity and purity of co-cultured myocardial cells, and （4）differentiation of MSCs into myocardial cells after co-culture. RESULTS： （1）Bone marrow cell suspension was inoculat

关 键 词：干细胞因子 骨髓细胞 间质干细胞 细胞分化 

分 类 号：R394.2[医药卫生—医学遗传学]