大鼠脂肪间充质干细胞的分离培养及生物学特性的实验研究

Experimental Study of the Cultrure,Isolation and Biological Chracteristics of Rat Adipose-derived Adult Stem Cells in Vitro

【目的】建立一种体外分离和培养大鼠脂肪间充质干细胞的方法,并探讨其部分生物学特性。【方法】无菌技术下切取双侧腹股沟脂肪垫,加入适量Ⅰ型胶原酶消化获取细胞,接种入含10%新生牛血清的DMEM培养液,倒置显微镜下连续观察细胞形态变化,测定其生长曲线及贴壁率。【结果】原代培养显示培养的脂肪间充质干细胞2 d左右开始贴壁,7 d左右可达90%融合,基本上为梭形成纤维样细胞形态。14代以前具有活跃的增殖能力。【结论】成功地建立了一种分离培养大鼠脂肪间充质干细胞的方法,其生长稳定、增殖较快,可以作为组织工程的种子细胞。

[ObjectivelTo establish a method for the isolation and culture of rat adipose-derived adult stem cells（ADAS） in vitro and explore their biological chracteristics. [Methods]The inguinal fat pads were excised with sterile technique, followed by digestion with 0. 075% collagenase, then the cells were cultured in Dulbecco modified Eagle medium with 10% fetal bovine serum. The morphology of ADAS was observed with inserted microscope constantly. Growth curve and adl^esiveness rate were detected. [Results]ADAS began to be adhesive after primary culture for 2 days. More than 90% ADAS fused at 7th day was basically fibroblast-shaped. ADAS showed active proliferative ability in passaging culture before 14th day. [Conclusion]The ioslation and culture of ADAS by the method successfully established in this study are stable growth and rapid proliferation in vitro. ADAS could be used as seed cells for tissue engineering.