药物诱导后胚胎胰腺干细胞胰岛素分泌研究

STUDY ON INSULIN SECRETION BY DRUG-INDUCED EMBRYONIC PANCREATIC STEM CELLS

目的探讨胚胎胰腺干细胞经药物诱导为胰岛素分泌细胞(IPCs)后的胰岛素分泌情况。方法体外分离培养获得的胚胎胰腺干细胞,经药物诱导后,双硫腙(DTZ)染色鉴定IPCs的形成情况;分别采用高浓度(25.0mmol/L)和低浓度(5.5mmol/L)的葡萄糖溶液刺激诱导后的细胞,ELISA方法检测该细胞群2周内胰岛素的分泌情况。结果经诱导后IPCs在两种浓度葡萄糖溶液刺激下均分泌胰岛素,且高浓度葡萄糖溶液刺激下胰岛素分泌量明显高于低浓度葡萄糖溶液(t=7.228,P<0.01)。IPCs在糖溶液刺激下第7天与第9天的胰岛素分泌量明显高于其他各天(F=38.373,q=3.033～6.085,P<0.05);第7天与第9天比较差异无显著性(P>0.05)。结论离体条件下,胚胎胰腺干细胞经药物诱导成为IPCs后,2周内可持续分泌胰岛素,其胰岛素释放量在诱导后第7～9天达到最大值,可作为胰岛移植的最佳时期。

Objective To explore the insulin secretion of insulin-producing cells （IPCs） derived from drug-induced em- bryonic stem cells （ESCs）. Methods Embryonic pancreatic stem cells were isolated and cultured in vitro, and induced by drug, the formation of IPCs was identified by dithizone （DTZ） staining. The induced cells were then stimulated by high-concentration （25.0 retool/L） and low-concentration glucose solution （5.5 mmol/L）, respectively. ELISA was applied to determine insulin pro- duction of the cells in two weeks. Results The induced IPCs secreted insulin after stimulation by low-or high-concentration glu- cose solution, and the cells stimulated by high-concentration one secreted much more insulin （t = 7.228, P 〈0.01）. The insulin se cretion of IPCs on days 7--9 after glucose stimulation was higher than other days （F=38.373 ,q=3.033--6.085, P〈0.05）. Con- clusion In vitro, IPCs derived from drug-induced ESCs can continuously secrete insulin for two weeks, and the highest level of in- sulin secretion was observed on days 7--9, which can be regarded as the best time for islet transplantation.