脂肪干细胞提取方法的改良.pdf

浙江临床医学2015年2月第17卷第2期 ·215· ·实验研究· 脂肪干细胞提取方法的改良 胡金灵  邵世飚  梁廷波 【摘要】 目的 探索一种简单高效的脂肪干细胞提取方法。方法 实验动物选取SD 大鼠，在传统脂肪干细胞提取方法的基础上提高胶原 酶浓度并省略裂解红细胞及筛网滤过步骤作改良，比较两种提取方法所用时间、提取的脂肪干细胞数量、生物学特性、传代及分化能力等。 结果 改良法提取脂肪干细胞用时70min ，提取5瓶原代细胞，未污染，细胞3d后传代，子代细胞生长良好，成功诱导分化为脂肪细胞。结 论 改良法提取脂肪干细胞能够显著增加细胞提取数量，缩短提取时间，降低原代细胞污染率，不影响细胞生物学特性及分化能力，是一种 简单高效的脂肪干细胞提取方法。 【关键词】 脂肪干细胞 改良 提取 【Abstract 】 Objective Explore a simple and efficient extraction method of adipose-derived stem cell. Methods A comparison was made between an improved method and the conventional method, the former increased concentration of collagenase and saved the steps of erythrocyte lysis and filtration by screen on the basis of traditional method. Adipose-derived stem cells were respectively isolated by traditional method and improved method. Extration time and the quatity ，biological characteristics and differentiation potential of adipose-derived stem cells were compared. Results the time for improved method was 70 minutes. There was no contamination in the five cell bottles containing a lot of impurity. The time needed to passage was 3 days and the cells passaged grew well and were induced to adipocyte successfully. While the time for traditional method was 128 minutes. There was contamination in one cells bottle out of three bottles containing a little of impurity. The time needed to passage was 5 days and the cells passaged grew well and were induced to adipocyte successfully. Conclusion The modified method can significantly increase the quality of the extracted cells， shorten the extracting time and decrease the contamination rate of primary cells ，while it would not affect the cell biological characteristics and the differentiation potential. So the method was simple and effective. 【Key word 】 Adipose-derived stem cells Improved Extract