小鼠3T3-L1前脂肪细胞培养与诱导分化方法的建立  

A novel approach for culture and induced differentiation of mouse 3T3-L1 preadipocytes

作　　者：郭秀玲 徐民岗 张秀丽 师磊[2] 陈显久[2] 

机构地区：[1]山西省长治市第二人民医院内分泌科,046000 [2]山西医科大学生物化学与分子生物学教研室

出　　处：《中国药物与临床》2013年第12期1542-1544,I0002,共4页Chinese Remedies & Clinics

基　　金：山西省国际科技合作项目(2012081050-1);山西省科技攻关项目(20110321076-02)

摘　　要：目的建立小鼠3T3-L1前脂肪细胞的培养及诱导分化为成熟脂肪细胞的方法。方法使用含10%胎牛血清（FBS）的高糖达氏修正伊氏培养基（DMEM）-F12液体培养基在体积分数5%二氧化碳（CO2）、37℃条件下常规培养3T3-L1细胞,2~3 d换液1次;诱导分化培养基Ⅰ培养2 d,诱导分化培养基Ⅱ培养2 d;基础培养基培养4~6 d,1~2 d换液1次。结果小鼠3T3-L1前脂肪细胞状态良好,成铺路石状生长,布满培养瓶底,3 d传代1次。90%以上细胞诱导分化成功,细胞呈圆形,有大量酯滴聚集,油红O染色呈橘红色。结论建立了小鼠3T3-L1前脂肪细胞的培养方法和诱导分化方法,为肥胖相关药物研究奠定了方法基础。Objective To establish the methodology for culture and induced differentiation for mouse 3T3-L1 preadipocytes. Methods The 3T3-L1 cells were incubated in glucose-rich DMEM/F12 medium containing 10% FBS under an atmosphere with 5% CO2 at 37 ℃, during which the incubation solution was replenished every 2 to 3 days. The induced differentiation followed the protocol of medium [ for 2 days, medium H for 2 clays and the basic medi- mn for 4-6 days, during which the incubation solution was replenished daily or every 2 clays intervals. Results Mouse 3T3-L1 preadipocytes grew in a paving stone fashion and appeared in good conditions. These cells covered the bottom of incubators, with the culture medium being replenished every 3 days. Differentiation was successfully achieved in over 90% of the cells, which were round-shaped, with massive orange ester droplets by oil-red O staining. Conclusion Our established methodology for culture and induced differentiation of mouse 3T3-L1 preadipocytes has laid the foun- dation for obesity-related drug researches.

关 键 词：3T3-L1细胞 细胞培养技术 细胞分化 小鼠 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]