Discussion on cultivation and methodology of four-drug combination-induced differentiation in mouse preadipocytes 3T3-L1 cells
10.11958/20160214
- VernacularTitle:小鼠前脂肪细胞3T3-L1培养与四联诱导分化方法的探讨
- Author:
Huizhi SUN
;
Derun TIAN
;
Jie MENG
;
Nan ZHAO
;
Jie HAN
;
Chunchun GAN
;
Yong WANG
- Publication Type:Journal Article
- Keywords:
adipocytes;
in vitro;
cell culture techniques;
3T3-L1 cells
- From:
Tianjin Medical Journal
2016;44(8):993-995
- CountryChina
- Language:Chinese
-
Abstract:
Objective To optimize and establish the methodology for culturing and inducing differentiation of mouse preadipocytes 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 mediumⅠin three-drug combination group including insulin 10 mg/L, IBMX 0.5 mmol/L and DEX 1.0μmol/L. The protocol of mediumⅠin four-drug combination group including indometacin 0.1 mmol/L based on those of three-drug combination group. Both of them were incubated for 2 days and continuous for 2 times. And medium Ⅱ included insulin 10 mg/L for 2-day culturing and continuous for 2 times. Oil red O staining was used to observe the morphological changes of two groups of cells before and after treatment under inverted microscope. Results Mouse preadipocytes 3T3-L1 appeared in good conditions and grew in a paving stone fashion. These cells covered homogeneously the bottom of incubators, the culture medium refreshed every 2 days. The results of four-drug combination group were better than those of three-drug combination group. After three-drug combination induced differentiation, there was no significant change in cell morphology. Comparing with three-drug combination 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 for culturing and inducing differentiation of mouse preadipocytes 3T3-L1 by adding indometacin on the basis of the three-drug combination induced differentiation.
