从人胎儿胰腺中分离出的巢蛋白阳性祖细胞具有与间充质干细胞相同的表型标志物。
Nestin-positive progenitor cells isolated from human fetal pancreas have phenotypic markers identical to mesenchymal stem cells.
作者信息
Zhang Ling, Hong Tian-Pei, Hu Jiang, Liu Yi-Nan, Wu Yong-Hua, Li Ling-Song
机构信息
Department of Endocrinology, Peking University Third Hospital, 49 Huayuanbeilu, Haidian District, Beijing 100083, China.
出版信息
World J Gastroenterol. 2005 May 21;11(19):2906-11. doi: 10.3748/wjg.v11.i19.2906.
AIM
To isolate nestin-positive progenitor cells from human fetal pancreas and to detect their surface markers and their capability of proliferation and differentiation into pancreatic islet endocrine cells in vitro.
METHODS
Islet-like cell clusters (ICCs) were isolated from human fetal pancreas by using collagenase digestion. The free-floating ICCs were handpicked and cultured in a new dish. After the ICCs developed into monolayer epithelium-like cells, they were passaged and induced for differentiation. Reverse transcription polymerase chain reaction (RT-PCR), immunofluorescence stain, fluorescence-activated cell sorting (FACS) and radioimmunoassay (RIA) were used to detect the expression of cell markers.
RESULTS
(1) The monolayer epithelium-like cells had highly proliferative potential and could be passaged more than 16 times in vitro; (2) RT-PCR analysis and immunofluorescence stain showed that these cells expressed both nestin and ABCG2, two of stem cell markers; (3) FACS analysis revealed that CD44, CD90 and CD147 were positive, whereas CD34, CD38, CD45, CD71, CD117, CD133 and HLA-DR were negative on the nestin-positive cells; (4) RT-PCR analysis showed that the mRNA expression of insulin, glucagon and pancreatic-duodenal homeobox gene-1 was detected, whereas the expression of nestin and neurogenin 3 disappeared in these cells treated with serum-free media supplemented with the cocktail of growth factors. Furthermore, the intra-cellular insulin content was detected by RIA after the induction culture.
CONCLUSION
Nestin-positive cells isolated from human fetal pancreas possess the characteristics of pancreatic progenitor cells since they have highly proliferative potential and the capability of differentiation into insulin-producing cells in vitro. Interestingly, the nestin-positive pancreatic progenitor cells share many phenotypic markers with mesenchymal stem cells derived from bone marrow.
目的
从人胎儿胰腺中分离巢蛋白阳性祖细胞，检测其表面标志物以及体外增殖和分化为胰岛内分泌细胞的能力。
方法
采用胶原酶消化法从人胎儿胰腺中分离胰岛样细胞团（ICCs）。挑选出游离的ICCs并接种于新培养皿中培养。待ICCs发育为单层上皮样细胞后传代并诱导分化。采用逆转录聚合酶链反应（RT-PCR）、免疫荧光染色、荧光激活细胞分选（FACS）和放射免疫分析（RIA）检测细胞标志物的表达。
结果
（1）单层上皮样细胞具有高度增殖潜能，可在体外传代16次以上；（2）RT-PCR分析和免疫荧光染色显示，这些细胞表达干细胞标志物巢蛋白和ABCG2；（3）FACS分析显示，巢蛋白阳性细胞上CD44、CD90和CD147呈阳性，而CD34、CD38、CD45、CD71、CD117、CD133和HLA-DR呈阴性；（4）RT-PCR分析显示，检测到胰岛素、胰高血糖素和胰十二指肠同源盒基因-1的mRNA表达，而在用添加生长因子混合物的无血清培养基处理的这些细胞中，巢蛋白和神经生成素3的表达消失。此外，诱导培养后通过RIA检测细胞内胰岛素含量。
结论
从人胎儿胰腺中分离出的巢蛋白阳性细胞具有胰腺祖细胞的特征，因为它们具有高度增殖潜能和体外分化为胰岛素分泌细胞的能力。有趣的是，巢蛋白阳性胰腺祖细胞与源自骨髓的间充质干细胞共享许多表型标志物。
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