【摘要】 目的改进小鼠前脂肪细胞3T3-L1的培养并诱导分化为成熟脂肪细胞的方法。方法使用含有10%胎牛血清(FBS)的高糖型DMEM液体培养基常规培养小鼠前脂肪细胞,2~3d换液1次。细胞按诱导分化方式的不同分为三联诱导组和四联诱导组。三联诱导组诱导分化培养基Ⅰ成分为常规培养基基础上加用人胰岛素10mg/L,1-甲基-3-异丁基黄嘌呤(IBMX)0.5mmol/L,地塞米松(DEX)1.0μmol/L;四联诱导组诱导分化培养基Ⅰ的成分为在三联诱导组基础上加入吲哚美辛0.1mmol/L。2组均诱导分化培养基Ⅰ培养2d,连续诱导2次后换用诱导分化培养基Ⅱ,成分为:高糖型DMEM培养基含10%胎牛血清、100U/m L青霉素和100mg/L链霉素混合液,人胰岛素10mg/L。培养2d,连续诱导2次。倒置显微镜和油红O染色观察诱导前及诱导后2组细胞的形态变化。结果小鼠前脂肪细胞3T3-L1状态良好,呈现铺路石状生长,均匀布满培养瓶底,2d传代1次。四联诱导组诱导分化结果优于三联诱导组。三联诱导剂诱导前脂肪细胞后,细胞形态并未发生明显变化。四联诱导剂诱导前脂肪细胞后,可达到90%以上的成熟脂肪细胞。成熟的脂肪细胞呈圆形,有大量脂滴聚集,油红O染色显现橘红色。结论在三联诱导基础上加入吲哚美辛的四联诱导法可以更好地促进脂肪前体细胞分化。更多还原

【Abstract】 Objective To optimize and establish the methodology for culturing and inducing differentiation of mousepreadipocytes 3T3-L1. Methods The mouse cells 3T3-L1 were incubated in DMEM medium contained with 10% FBS,during which the incubation medium was refreshed every 2 to 3 days. Two methods were used to introduce differentiation,including three- drug combination group and four- drug combination group. The protocol of medium Ⅰ in three- drugcombination group including insulin 10mg/L, IBMX 0.5mmol/L and DEX 1.0μmol/L. The protocol of mediumⅠin four-drug combination group including indometacin 0.1mmol/L based on those of three-drug combination group. Both of themwere incubated for 2 days and continuous for 2 times. And medium Ⅱ included insulin 10mg/L for 2-day culturing andcontinuous for 2 times. Oil red O staining was used to observe the morphological changes of two groups of cells before andafter treatment under inverted microscope. Results Mouse preadipocytes 3T3-L1 appeared in good conditions and grew ina paving stone fashion. These cells covered homogeneously the bottom of incubators, the culture medium refreshed every 2days. The results of four-drug combination group were better than those of three-drug combination group. After three-drugcombination induced differentiation, there was no significant change in cell morphology. Comparing with three- drugcombination induced differentiation, four- drug combination was successfully achieved in over 90% of the cell inducing,which were round-shaped, with jacinth ester droplets by oil-red O staining. Conclusion We have optimized the method forculturing and inducing differentiation of mouse preadipocytes 3T3-L1 by adding indometacin on the basis of the three-drugcombination induced differentiation.更多还原