膀胱匀浆上清液在大鼠骨髓间充质干细胞诱导分化为平滑肌样细胞中的作用  

Rat bone marrow mesenchymal stem cells differentiate into smooth muscle cells in vitro induced by the supernatant of homogenized bladders

作　　者：王振显[1] 蔡文清[1] 宋永周[2] 瞿长宝[1] 史正亮[2] 郭威[3] 

机构地区：[1]河北医科大学第二医院泌尿外科,河北省石家庄市050051 [2]河北医科大学第二医院骨科,河北省石家庄市050051 [3]河北医科大学解剖教研室,河北省石家庄市050017

出　　处：《中国组织工程研究与临床康复》2008年第8期1422-1425,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：目的：环境因素对干细胞的分化起到重要的作用。实验拟观察经低浓度二甲亚枫进行预诱导后，大鼠骨髓间充质干细胞在体外经膀胱组织匀浆上清液诱导定向分化为平滑肌样细胞的可行性。方法：实验于2007-04／2007-09在河北医科大学解剖教研室下属细胞培养室完成。①实验材料：4～6周龄100～150g健康清洁级SD大鼠1只，雌雄不限，8周龄220～250gSD大鼠4只，由河北医科大学实验动物中心提供，实验过程中对动物处置符合动物伦理学标准。②实验方法：采用骨髓干细胞培养基和贴壁法从SD大鼠骨髓中分离骨髓间充质干细胞，并在体外扩增、传代。选择8周龄SD大鼠进行膀胱组织上清液的制备。取第4代骨髓间充质干细胞用1％二甲亚砜预诱导8h后，应用膀胱匀浆上清液诱导7d。以未进行诱导的骨髓间充质干细胞为阴性对照组。③实验评估：观察细胞形态学的变化，并运用免疫细胞化学检测特异性。一平滑肌肌动蛋白（α—SMA）的表达。结果：骨髓间充质干细胞经诱导分化后，细胞呈梭形平滑肌样，融合后形成峰谷状排列，并表达平滑肌特异性蛋白标志物。一平滑肌肌动蛋白，诱导分化率为（45．6±3.5）％，与阴性对照组相比差异具有显著性意义（P〈0．05）。结论：采用此方法成功诱导骨髓间充质干细胞分化为平滑肌样细胞。AIM： Environmental factors play important roles in the differentiation of stem cells. This study explored the potential of rat bone marrow mesenchymal stem cells （MSCs） to differentiate into smooth muscle cells by the supernatant of homogenized bladder. METHODS： The experiment was conducted in the cell culture laboratory affiliated to the Department of Anatomy, Hebei Medical University from April to September 2007. ①One 4-week-old healthy SD rat of 100-150 g, either male or female, and four 8-week-old SD rats of 221-250 g were provided by the experimental animal center of Hebei Medical University. All procedures employed in this study were consistent with the animal ethical standards. ②MSCs were isolated from SD rat bone marrow by total marrow culture associated with attachment method, purified and cultured in vitro. The supernatant of homogenized bladder was made with 8-week-old SD rats. The MSCs of the 4^th generation were cultured in DMEM with 1% dimethyl sulfoxide （DMSO） for 8 hours before changing the supernatant of homogenized bladders to induce differentiation for 7 days. MSCs not treated served as negative control. ③Morphology of cells was observed and immunocytochemistry was performed to detect the expression of specific α-smooth muscle actin （SMA）. RESULTS： After induction, MSCs showed smooth muscle-like cells and spindle-shaped. The cells fused as a hill-and-valley growth pattern, and expressed the specific a -SMA. The differentiation rate was （45.6±3.5）%, which was stati.stically significant compared with negative control group （P 〈 0.05）. CONCLUSION： MSCs are induced into smooth muscle cells in vitro by this method.

关 键 词：干细胞 平滑肌 细胞培养 分化 

分 类 号：R318.4[医药卫生—生物医学工程]