小鼠附睾脂肪干细胞分离培养及鉴定.docxVIP

小鼠附睾脂肪干细胞的分离培养及鉴定张鉴清1，季佳霖2，崔新明2，张? 祺3，李艳茹21吉林大学第四医院，吉林省长春市? 130011；2吉林大学病理生物学教育部重点实验室，吉林大学基础医学院病理学系，吉林省长春市? 130021；3吉林大学，吉林省长春市? 130021Isolation, culture and identification of adipose-derived stem cells from mouse epididymis?Zhang Jian-qing1, Ji Jia-lin2, Cui Xin-ming2, Zhang Qi3, Li Yan-ru21 the Fourth Hospital of Jilin University, Changchun 130011, Jilin Province, China; 2 the Key Laboratory of Pathobiology of Ministry of Education, Department of Pathology, School of Basic Medical Sciences, Jilin University, Changchun 130021, Jilin Province, China; 3 Jilin University, Changchun 130021, Jilin Province, China摘要? 背景：脂肪干细胞作为一种新的成体干细胞，具有来源丰富、取材容易、增殖能力强等优点，受到人们越来越多地关注。目的：体外分离培养小鼠附睾脂肪组织中的脂肪干细胞，并鉴定其生物学特性。方法：无菌切取小鼠附睾处脂肪组织，采用胶原酶进行消化，利用1次消化多次收集与差速贴壁法分离纯化脂肪干细胞。倒置显微镜及透射电子显微镜观察脂肪干细胞的形态。绘制脂肪干细胞的生长曲线。流式细胞术检测脂肪干细胞的免疫表型。加入细胞诱导剂对脂肪干细胞进行成骨及成脂肪的诱导分化。扫描电子显微镜观察脂肪干细胞与胶原支架材料的相容性。结果与结论：倒置显微镜下可见脂肪干细胞呈长梭形或成纤维细胞样，密集排列成漩涡状或成束状交织，可在体外稳定传至第9代。透射电子显微镜下可见脂肪干细胞表面有丰富的微绒毛结构，细胞核体积大，细胞质内可见线粒体、粗面内质网等细胞器。脂肪干细胞表达CD44、CD29，不表达CD34。经成脂肪诱导剂诱导后，多数脂肪干细胞胞质内可见小脂滴，油红O染色可将小脂滴染成红色。经成骨诱导剂诱导后，在脂肪干细胞生长密集区域内的细胞结构模糊、细胞间界限不清楚。经茜素红染色后，可见较多大小不一的折光率较强颗粒状结构沉积。扫描电子显微镜观察到脂肪干细胞呈铺展状生长于胶原支架材料上。结果表明该方法分离出的脂肪干细胞能在体外扩增并稳定传代，在一定诱导条件下，能够分化为成骨细胞和脂肪细胞并且与胶原支架具有良好的相容性。中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程全文链接：窗体顶端关键词：干细胞；脂肪干细胞；小鼠；附睾；脂肪组织；差速贴壁；培养； ?? Abstract：BACKGROUND: As a new kind of adult stem cells, adipose-derived stem cells get more and more attention, because of rich source, drawing materials easily and powerful proliferation.? OBJECTIVE: To isolate and culture adipose-derived stem cells from the epididymal adipose tissue in mice, and to identify their biological characteristics.METHODS: Adipose tissue was obtained from epididymis in mice by aseptically cutting. The tissue was digested using collagenase. Adipose-derived stem cells were separated and purified by using one digestion, multiple collection method and differential adhesion method. The morphology of adipose-derived stem cells was obs