摘要
背景:有文献证实以酶消化法可获得在脂肪组织间充质干细胞,传至第10代时细胞生长仍良好,但表面标志物表达下降。目的:分离培养大鼠脂肪组织间充质干细胞,传代至第4代,检测免疫表型。方法:从6只SD大鼠腹股沟脂肪垫中分离培养脂肪组织间充质干细胞,以贴壁细胞扩增传代至第4代。相差显微镜下观察脂肪组织间充质干细胞的形态,并用流式细胞术鉴定脂肪组织间充质干细胞的表面标志物CD11b,CD29,CD45,CD49d,CD90和CD106的表达率。结果与结论:大鼠脂肪组织间充质干细胞呈现类似成纤维细胞样形态学特征,免疫表型分析显示CD29和CD90呈阳性,而CD11b、CD45、CD49d和CD106呈阴性反应。结果提示通过这种酶消化法获得的SD大鼠的脂肪组织间充质干细胞,免疫表型分析显示符合间充质干细胞的特征。
BACKGROUND:Many studies have confirmed that adipose tissue-derived mesenchymal stem cells (ADMSCs) can be collected using enzyme digestion method.At passage 10,ADMSCs grew well,but their surface marker expression decreases.OBJECTIVE:To isolate and cultivate rat ADMSCs and detect immunophenotype at passage 4.METHODS:ADMSCs of six Sprague-Dawley rats from inguinal fat pads were isolated and cultured.Adherent cells were amplified till passage 4.Morphology of ADMSCs was observed under a phase contrast microscope.The immunophenotype markers including CD11b,CD29,CD45,CD49d,CD90 and CD106 of ADMSCs were determined using flow cytometry.RESULTS AND CONCLUSION:Rat ADMSCs showed fibroblastic-like morphologic characteristics.The immunophenotype analysis revealed that the rat ADMSCs were positive for CD29 and CD90,but negative for CD11b,CD45,CD49d and CD106.The rat ADMSCs can be obtained through enzyme-based isolation procedures.The results revealed that the immunophenotype characterization of rat ADMSCs is consistent with mesenchymal stem cells.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2010年第23期4207-4210,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
国家重点基础研究计划(973计划)项目(2007CB512705)
国家自然科学基金青年科学基金项目(30801464)资助~~