CAJ | 학술논문

目的探讨不同浓度重组人碱性成纤维细胞生长因子(recombinant human basic fibro-blast growth factor,rh-bFGF)对体外培养的人脂肪来源干细胞(human adipose-derived stem cells,hASCs)诱导分化为脂肪细胞的影响,通过实验寻找促进hASCs向脂肪细胞转化的最佳浓度.方法取健康脂肪抽吸者脂肪混悬液进行分离提取hASCs,进行hASCs的培养、鉴定和成脂诱导分化,并在成脂诱导的细胞中分别加入0 ng/ml rh-bFGF作为对照组,加入10、20、40 ng/ml外源性rh-bFGF作为实验组.MTT比色法检测成脂细胞的增殖速率,油红O染色定性分析新形成脂肪细胞的时间,利用Western印迹法检测成熟脂滴表面标记蛋白CIDEC的表达.结果添加40ng/mlrh-bFGF的脂滴形成平均时间为(11.5±1.9)h,检测细胞增殖的吸光度值平均为0.52±0.10,10、20、40 ng/mlrh-bFGF对细胞的增殖均有促进作用,尤以40ng/ml浓度最为明显,40ng/mlrh-bFGF成熟脂滴表面蛋白CIDEC的表达量高于其他组.结论在hASCs成脂诱导剂中添加rh-bFGF能有效促进成脂细胞的增殖速率,加速hASCs向脂肪细胞的分化,其中以40ng/ml rh-bFGF为最佳浓度.
목적 탐토불동농도중조인감성성섬유세포생장인자(recombinant human basic fibro-blast growth factor,rh-bFGF)대체외배양적인지방래원간세포(human adipose-derived stem cells,hASCs)유도분화위지방세포적영향,통과실험심조촉진hASCs향지방세포전화적최가농도.방법 취건강지방추흡자지방혼현액진행분리제취hASCs,진행hASCs적배양、감정화성지유도분화,병재성지유도적세포중분별가입0 ng/ml rh-bFGF작위대조조,가입10、20、40 ng/ml외원성rh-bFGF작위실험조.MTT비색법검측성지세포적증식속솔,유홍O염색정성분석신형성지방세포적시간,이용Western인적법검측성숙지적표면표기단백CIDEC적표체.결과 첨가40ng/mlrh-bFGF적지적형성평균시간위(11.5±1.9)h,검측세포증식적흡광도치평균위0.52±0.10,10、20、40 ng/mlrh-bFGF대세포적증식균유촉진작용,우이40ng/ml농도최위명현,40ng/mlrh-bFGF성숙지적표면단백CIDEC적표체량고우기타조.결론 재hASCs성지유도제중첨가rh-bFGF능유효촉진성지세포적증식속솔,가속hASCs향지방세포적분화,기중이40ng/ml rh-bFGF위최가농도.
Objective To study the effect of the exogenous recombinant human FGF-basic with different concentrations upon inducing human adipose-derived stem cells (hASCs) to differentiate into adipose cells,and the optimum concentration of exogenous rh-bFGF by experimental research.Methods hASCs were isolated and extracted by enzymatic digestion from the liposuction aspirate.hASCs using adipogenic supplement were divided into experimental group and blank group:the experimental group of adipogenic supplement was divided into adding the exogenous rh-bFGF 10 ng/ml,20 ng/ml and 40 ng/ml,the blank group of adipogenic supplement was cultured without exogenous rh-bFGF.MTT method was used to detect the adipocytes proliferation.The oil red O staining was used in the qualitative analysis on the time of newly forming adipocyte cells.Western blot was used to detect the effects of rh-bFGF on the expression of lipid droplets surface protein CIDEC at different stages during the culture.Results The experimental group could obviously shorten the period of inducing hASCs to differentiate into adioicytes,and promote the proliferation of adipocytes.The formation rate and the proliferation of adipocytes in the group adding 40 ng/ml rh-bFGF were superior to those in the experimental group else and blank group.The average time of the newly formed lipid droplets by adding 40 ng/ml rh-bFGFwas (11.5±1.9)h.The average absorbance of cell proliferation by adding 40ml rh-bFGF was 0.52 ±0.10.The CIDEC expression quantity of adding 40 ng/ml rh-bFGF group was also superior to that in the experimental group and blank group.Conclusions rh-bFGF in hASCs adipogenic supplement could promote the proliferation of adipocytes and dramatically accelerates the program of hASCs differentiating to adipocytes,in which the optimum concentration of rh-bFGF is 40ng/ml.