大鼠前脂肪细胞消化酶原代培养方法改良的探讨  

Modification of collagen digestion method on isolation and culture of rat preadipocyte

作　　者：蒋茂莹[1] 张艳[1] 刘友学[1] 

机构地区：[1]重庆医科大学附属儿童医院儿童营养研究中心,重庆400014

出　　处：《重庆医科大学学报》2009年第6期743-746,共4页Journal of Chongqing Medical University

基　　金：国家自然科学基金资助项目(基金编号:30571565)

摘　　要：目的:改进大鼠前脂肪细胞的原代培养方法。方法:对Poznanski胶原酶消化法进行改良,包括延长胶原酶消化法时间、3次离心和2次滤网过滤,分化培养后采用油红O染色对细胞进行鉴定,通过形态学观察、生长曲线的绘制确定原代培养大鼠前脂肪细胞的细胞学特性。结果:用改良后的方法获得大量成份均一、生长旺盛的前脂肪细胞,细胞活力测定90%为活细胞;前脂肪细胞5d左右进入指数增长期,8d左右达到平台期,倍增时间约为60h;在分化培养基的诱导下可以向脂肪细胞分化。结论:实验成功地建立了前脂肪细胞培养体系,较原方法更加有效。Objective： To establish a better method for isolating and culturing preadipocytes from rat white adipose tissue. Methods： The adipose tissue was digested by collagen I and preadipocytes were collected by centrifugations and filtrations. Cells were cultured using basic and differentiation media. Preadipocytes were detected by morphologic method, auxodrome study and stained with Oil red O for determining the cell identity. Results： Using our modified method, primary preadipocytes can be harvested in greater amount compared with the original method. The primary preadipocytes could differentiate into adipocytes under the induction of differentiation medium. The preadipocytes obtained by our method had the characteristics of preadipocytes. Conclusion： We have established an efficient preadipocyte culture method. Our medication method is better than the original method.

关 键 词：大鼠 前脂肪细胞 培养方法 改良 

分 类 号：R72[医药卫生—儿科]