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成人胰腺干细胞转分化为胰岛的研究
作者姓名:Song Z  Gu K
作者单位:1. 710032,西安,第四军医大学西京医院肝胆外科
2. 710032,西安,第四军医大学西京医院感染管理科
摘    要:目的:通过对成人胰腺干细胞转分化为胰岛过程的研究以便更深入了解及改进胰腺干细胞分离、培养、鉴定方法。方法:成人胰腺组织经胶原酶消化后,用密度梯度离心法将胰腺外分泌细胞、导管上皮细胞和胰岛分离、纯化,导管上皮细胞即具有转分化潜能的干细胞,在体外先后以CMRL1066和无血清DMEM/F12培养液共培养27d,在培养的不同时间点取样本于光镜和电镜下观察细胞形态学变化及干细胞特异性转录基因PDX-1,CK-19蛋白等单抗的免疫组化染色,并测定培养液中的淀粉酶和胰岛素含量。结果:上述方法可获得大量以往在胰岛分离时丢弃的胰腺导管上皮细胞。经体外一定条件的培养后,第1天即可见PDX-1,CK-19阳性细胞,胰腺导管上皮细胞迅速分裂增殖并转变为有分化能力的干细胞继而转分化为三维结构的胰岛细胞。培养27d后,平均每克胰腺组织可生成760个胰岛。结论:成人胰腺的导管上皮具有干细胞潜能并可在体外转分化为大量具有内分泌功能的胰岛,用此方法获得大量的胰岛可能为克服胰岛移植的供体短缺提供一条新的途径。

关 键 词:胰腺  干细胞  胰岛  转分化  糖尿病  同种移植

Transdifferentiating of adult human pancreatic stem cells into islet cells
Song Z,Gu K.Transdifferentiating of adult human pancreatic stem cells into islet cells[J].Chinese Journal of Surgery,2002,40(11):807-810,W001.
Authors:Song Zhenshun  Gu Keju
Institution:Department of Hepato-Biliary Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China.
Abstract:Objective To understand and improve the transdifferentiation of pancreatic stem cells into islet cells through isolation, cultivation and transdifferentiation of adult human pancreatic duct cells. Methods A portion of adult human pancreas was digested with collagenase, followed by incontinuous density gradient to separate islets from the acinar and ductal tissue. Duct epithelial cells were cultivated in CMRL1066 and then in serum free DMEM/F12 medium with the addition of growth factors for 27 days. Samples were taken at different time points for light and electron microscopic examination and for immunocytochemical study with antibodies against transdifferentiation gene PDX 1 and protein CK 19. Amylase and insulin contents in the medium were assayed. Results A large number of duct epithelial cells were harvested after the isolation of islets. Some duct epithelial cells were PDX 1 and CK 19 positive at day one and duct epithelial cells proliferated and expanded rapidly and then transdifferentiated into stem cells and finally 3D islets. 760 islets were harvested from each gram of pancreatic tissue on day 27. Conclusions Adult pancreatic duct cells are potential stem cells and could be transdifferentiated into islet cells in vitro under appropriate conditions.
Keywords:Pancreas  Stem cells  Islets of langerhans  Transdifferentiation
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