小鼠胰腺干细胞的分离、培养与鉴定  

Isolation,culture and identification of pancreatic stem cells from newborn mouse

作　　者：李军[1] 孙毅[2] 马荣[2] 李勇[2] 

机构地区：[1]宁夏职业技术学院生物与制药技术系,银川750021 [2]宁夏大学生命科学学院西部特色动植物资源保护与利用教育部重点实验室

出　　处：《山东医药》2012年第41期29-31,共3页Shandong Medical Journal

基　　金：宁夏回族自治区自然科学基金资助项目(NZ1161;NZ0905)

摘　　要：目的从新生小鼠中分离培养并鉴定胰腺干细胞。方法利用组织块法分离新生小鼠胰腺源性干细胞,经RPMI1640+5%胎牛血清+双抗+1 mmol/L丙酮酸钠+1 mmol/L非必需氨基酸+10 ng/mL表皮生长因子+10 ng/mL成纤维生长因子+2 mmol/L谷氨酰胺的溶液培养,利用免疫组化分析胰腺源性干细胞的兔抗巢蛋白抗体(Nestin)、胰肠同源域因子(PDX-1)和葡葡糖转运子(GLUT-2)抗原表达;经10 mmol/L尼克酰胺诱导产生胰岛样细胞团并利用双硫腙(DTZ)染色分析其胰岛素分泌情况。结果分离培养的胰腺源性干细胞呈Nestin、PDX-1和GLUT-2阳性;诱导的胰岛样细胞团呈DTZ染色阳性。结论分离获得的新生小鼠胰腺源性干细胞在体外可分化为类胰岛,并具有胰岛素分泌功能。Objective To separate, culture and identify the pancreatic stem cells （PSCs） from newborn mouse. Methods The pancreas-derived stem cells were separated from newborn mouse by tissue block method and euhured in the liquid medium （RPMI1640 ＋ 5% FBS ＋ double-antibiotics ＋ 1 mmol/L sodium pyrnvate ＋ 1 mmol/L Non-essential amino acid ＋ 10 ng/mL EGF ＋ 10 ng/mL bFGF ＋ 2 mmol/L Glutamine）. The expressions of Nestin, PDX-1 and GLUT-2 anti- gens were identified and analyzed by immunohistoehemistry. The pancreas-derived stem cells were induced by 10 mmol/L NIC and differentiated into panereatie islet-like eell clusters, which was stained with DTZ for analyzing the seeretion of in- sulin. Results In the isolated and cultured pancreas-derived stem cells, Nestin, PDX-1 and GLUT-2 expressed positive ly, in the induced panereatie islet-like cell elusters, DTZ expressed positively. Conclusions The pancreas-derived stem cells isolated from newborn mouse are PSCs, which can differentiate into similar islets of pancreas, and have islets of pan- ereas function.

关 键 词：胰腺干细胞 分离 培养 小鼠 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]