GLP-1、Betacellulin、Activin A、bFGF和Nicotinamide联合诱导小鼠胚胎干细胞分化为胰岛素分泌细胞  

Differentiation of Mouse Embryonic Stem Cells into Insulin-secreting Cells Induced by GLP-1,Betacellulin,Activin A,bFGF and Nicotinamide

作　　者：吴木潮[1] 陈黎红[1] 徐明彤[1] 程桦[1] 

机构地区：[1]中山大学附属第二医院内分泌科,广东广州510120

出　　处：《中山大学学报（医学科学版）》2004年第5期426-430,共5页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：中山大学附属第二医院重点扶持专科基金资助项目(F002002003)

摘　　要：【目的】探讨体外联合应用GLP-1、betacellulin、activinA、bFGF和nicotinamide5种生长因子能否将小鼠胚胎干细胞(ES细胞)诱导分化为胰岛素分泌细胞。【方法】以GLP-1、betacellulin、activinA、bFGF和nicoti-namide5种生长因子诱导E14.1小鼠ES细胞30d后,应用RT-PCR、DTZ染色和免疫组化检测分化细胞胰岛素表达,以流式细胞仪检测胰岛素分泌细胞的分化率,用RIA法测定培养液中胰岛素水平。【结果】分化细胞可检测到胰岛素mRNA表达,DTZ染色和胰岛素免疫组化染色阳性,胰岛素分泌细胞的分化率平均为13.6%±3.7%,在5.6mmol/L和25mmol/L葡萄糖浓度作用下培养液中胰岛素水平分别为(0.04±0.01)ng/mL和(0.09±0.02)ng/mL。【结论】体外联合应用GLP-1、betacellulin、activinA、bFGF和nicotinamide5种生长因子能将小鼠ES细胞诱导分化为胰岛素分泌细胞,但胰岛素分泌功能较差。To induce mouse embryonic stem cells(ES cells)to differentiate into insulin-secreting cells by GLP-1,b etacellulin,activin A,basic fibro blast growth factor(bFGF)and nicotinamide in vitro.E14.1mouse ES cells were treated by g lucagon-like peptide 1(GLP-1),betacellulin,activin A,bFGF,and nicotinamide for 30days,then insul in expression was examined by reverse transcription p olymerase chain reaction(RT-PCR),DTZ-staining,and immunohistochemistry.The percent age of insulin-secreting cells in di fferentiated cells were evaluated by flow cytometry and insulin level i n the medium was measured by RIA.The mRNA for insulin was detected in the differen tiated cells.DTZ-staining and immu nohistochemical staining for insulin were observed in the parts of the differentiated cells.13.6%±3.7%of the differentiated cells was insulin-secreting cells.In the presence of 5.6mmol /L and 25mmol /L glucose,the insulin levels in the medium were(0.04±0.01)ng /mL and(0.09±0.02)ng /mL,respectively.[Conclusion]Mouse ES cells can be induced to diffe rentiate into insulin-secreting ce lls by GLP-1,betacellulin,activin A,bFGF and ni cotinamide in vitro,however,the function of insulin-s ecreting cells is rather poor.

关 键 词：胰岛素分泌细胞 GLP-1 FGF 生长因子 胰岛素水平 干细胞分化 联合诱导 分化细胞 小鼠胚胎 ES细胞 

分 类 号：R587.1[医药卫生—内分泌] R335.6[医药卫生—内科学]