树大网膜前脂肪细胞原代培养及诱导分化

Primary culture and induction differentiation of Tupaia belangeri omental preadipocytes

目的探讨树大网膜前脂肪细胞原代培养的方法,优化其诱导分化的方法。方法采用酶消化的方法对树大网膜组织进行原代培养,待其传至第三代时,用含0.5mmoL/L3-异丁基-1-甲基黄嘌呤、10μmol/L胰岛素、0.2mmoL/L吲哚美辛、1μmoL/L地塞米松的高糖DMEM诱导配方对树大网膜前脂肪细胞进行成脂诱导,14d后见到细胞内聚集脂滴,对其进行油红O染色。结果树大网膜脂肪细胞在体外成功培养,细胞形态良好,呈梭形。诱导后的脂肪细胞第3d出现少数出现小脂滴,14d左右,大部分细胞胞质中出现圆而大的脂滴。结论树大网膜脂肪组织可在体外进行培养,稳定传代并能诱导分化为成熟的脂肪细胞。

Objective To study the primary culture method of Tupaia belangeri omental preadipocytes,and optimize the ways of induction differentiation.Methods Spindle cells from the Tupaia belangeri omental adipose tissues were cultured by enzyme-digesting method.Following complete confluence of the third passage of omental preadipocytes,differentiation was induced by HDMEM supplemented with 0.5 mmol/L 3-isobutyl-1-methylxanthine, 0.2mmol/L indomethacin,1μmol/L dexamethasone,and 10μmol/L insulin.After the 14th day,lipid droplets appeared in adipocytes,and were identified by oil red O staining.Results Tupaia belangeri omental preadipocytes were successfully cultured in vitro,cell shape was well and displayed into the spindle.Induction on the third day there appeared a few small lipid droplets in adipocytes.After the 14th days,most of adipocytes were filled with round and large lipid droplets.Conclusion Tupaia belangeri omental preadipocytes are successfully cultured in vitro,which can be induced into mature adipocyte under appropriate stimulus.