仔猪胰腺干细胞的多向分化潜能  

Multiple differentiation potentialities of pancreatic stem cells in neonatal piglets

作　　者：张慧茹[1,2] 冯若鹏 庞全海[2,3] 刘珍[1] 窦忠英 

机构地区：[1]河南工业大学生物工程学院,河南郑州450001 [2]国家干细胞工程技术研究中心陕西省分中心,陕西杨凌712100 [3]山西农业大学动物科技学院,山西太谷030801

出　　处：《中国兽医科学》2011年第1期70-74,共5页Chinese Veterinary Science

基　　金：国家高技术研究发展计划(863)项目(2005AA219050);国家自然科学基金资助项目(30671067);河南工业大学人才引进项目(150214)

摘　　要：以分离自1日龄仔猪体内的胰腺干细胞为检测细胞,诱导其向胰腺β细胞和三胚层功能性细胞分化,以证实分离的胰腺干细胞具有多向分化潜能。采用RPMI 1640培养液添加烟碱和肝细胞生长因子诱导胰腺干细胞向β细胞分化;添加5-氮胞苷向心肌细胞诱导;用地塞米松、β-磷酸甘油和抗坏血酸盐联合向成骨细胞诱导;用不同浓度的β-巯基乙醇作用不同时间向神经细胞诱导。结果表明,分离的仔猪胰腺干细胞在向胰腺β细胞诱导后,表现出胰岛素特征性双硫腙染色阳性,不同浓度的葡萄糖刺激诱导胰腺干细胞可分泌一定量的胰岛素;向心肌细胞诱导后,表现心肌细胞的α-actin和肌糖原阳性;向成骨细胞诱导后,细胞表现成骨细胞的碱性磷酸酶阳性和基质细胞钙离子茜素红染色阳性;向神经细胞诱导后,表达神经细胞的神经胶质纤维酸性蛋白、髓磷脂碱性蛋白、神经丝蛋白-200等多种表面标志。证实分离的仔猪胰腺干细胞具有向三胚层功能性细胞分化的多向分化潜能。The pancreatic stem cells,isolated from 1-day-old neonatal piglets,would be proved that they had the multipotent potentiality of stem cells by the inducing tests toward the function cells of pancreatic and triploblastic.The RPMI 1640 medium supplemented nicotinamide and hepatic growth factor was the differentiating medium toward endocrine β-cell of islets.The differentiating medium toward myocardium contained 5-Azacitidine.The RPMI 1640 medium associated with dexamethasone,β-glycerophosphate and ascorbic acid differentiated the cells toward the osteoblast lineage.To differentiate toward neural cells,the medium was supplemented different concentration β-mercaptoethanol.The results demonstrated that the isolated pancreatic stem cells were induced the differentiation toward endocrine β-cell,and stained dithizone-positive.The quantity of the secreted insulin varied with the different concentration of glucose.The pancreatic stem cells of differentiating toward myocardium expressed α-actin myocardial protein,and presented positive reaction of periodic acid Schiff.The isolated cells toward the osteoblast lineage were stained typical bright red with Alizarin Red and alkaline phosphatase.The induced cells expressed many markers of neural cells,which included GAFP,BMP and NF-200.In conclusion,the isolated stem cells from pancreas have the multipotent potentiality differentiating toward triploblastic function-cells in neonatal piglets.

关 键 词：胰腺干细胞 仔猪 多向分化潜能 诱导 

分 类 号：Q813.11[生物学—生物工程]