CAJ | 학술논문

目的：观察胎鼠胰腺干细胞体外对胰岛功能的保护作用。方法将孕16 d的SD大鼠胎鼠胰腺干细胞分离、纯化、培养传代，免疫细胞化学法及流式细胞术鉴定；分离纯化SD大鼠胰岛，双硫腙鉴定后分为A组和B组，分别行单纯胰岛培养及胰岛与胰腺干细胞联合培养14 d，期间观察胰岛形态变化、检测胰岛存活率及细胞凋亡率， ELISA法检测胰岛素分泌量、计算刺激指数。取两组培养7 d后悬浮生长的胰岛移植入糖尿病大鼠左肾包膜下，术后每天尾静脉采血以快速血糖测试仪检测血糖。结果胎鼠胰腺干细胞培养传代3代后免疫细胞化学可见巢蛋白（Nestin）阳性细胞，流式细胞术测定其含量占74．1％；培养第7、14天时B组胰岛存活率显著高于A组（P均＜0．01），培养第7天时B组胰岛细胞凋亡率显著低于A组（P＜0．05）；培养第7、14天时B组高糖刺激后胰岛素分泌量及刺激指数均显著高于A组（P均＜0．01）；B组移植后大鼠血糖第5天降至正常。结论胎鼠胰腺干细胞与胰岛联合培养可明显延长胰岛体外存活时间并使其保持良好的活性。
목적：관찰태서이선간세포체외대이도공능적보호작용。방법장잉16 d적SD대서태서이선간세포분리、순화、배양전대，면역세포화학법급류식세포술감정；분리순화SD대서이도，쌍류종감정후분위A조화B조，분별행단순이도배양급이도여이선간세포연합배양14 d，기간관찰이도형태변화、검측이도존활솔급세포조망솔， ELISA법검측이도소분비량、계산자격지수。취량조배양7 d후현부생장적이도이식입당뇨병대서좌신포막하，술후매천미정맥채혈이쾌속혈당측시의검측혈당。결과태서이선간세포배양전대3대후면역세포화학가견소단백（Nestin）양성세포，류식세포술측정기함량점74．1％；배양제7、14천시B조이도존활솔현저고우A조（P균＜0．01），배양제7천시B조이도세포조망솔현저저우A조（P＜0．05）；배양제7、14천시B조고당자격후이도소분비량급자격지수균현저고우A조（P균＜0．01）；B조이식후대서혈당제5천강지정상。결론태서이선간세포여이도연합배양가명현연장이도체외존활시간병사기보지량호적활성。
Objective To investigate the protective effect of pancreatic stem cells ( PSCs) of fetal rats in vitro on the islets of langerhans.Methods The PSCs of SD rat embryos on embryonic 16th day were isolated, purified, cultivated and passaged , then were identified by immuocytochemistry and flow cytometry .The rat's islets were isolated from the pancreas . After being identified by dithizone , the islets were divided into two groups:group A which underwent islet alone culturing and group B which underwent co-culturing of islet and PSCs for 14 days.The morphological changes , survival rate and ap-optosis rate of islets were observed under inverted microscope , fluorescence microscope and flow cytometry ( FCM) in both groups.The amount of insulin secretion was measured by ELISA;and insulin stimulation index was calculated with the in-sulin release test.After being cultured for 7 days, the islets in the two groups were transplanted to the peplos of the left kid-ney in SD rats with diabetes , respectively .The blood glucose of the SD rats was tested after transplantation .Results The immuocytochemistry showed the PSCs were nestin-positive after being cultivated and passaged for the 3rd generation.Flow cytometry showed its content accounted for 74.1%;on the 7th, 14th day, the survival rate of islets in group B was signifi-cantly higher than that in group A (all P<0.01), while on the 7th day, the apoptosis rate of islets in group B was signifi-cantly lower than that in group A (P<0.05).On the 7th , 14th day, under the high glucose stimulation , the levels of in-sulin secretion and insulin stimulation index in group B were significantly higher than those in group A (all P<0.01).Af-ter transplantation in group B , the blood glucose of rats dropped to normal on the 5th day.Conclusion Co-cultivation of islet and PSCs can significantly prolong the survival time of islets and preserve viability of islets .