胎儿胰腺来源的单克隆SP细胞具有间充质的表型特征  

Monoclonal SP cells derived from human fetal pancreas have phenotypic markers identical to mesenchymal stem cells

作　　者：吴永华[1] 洪天配[1] 张玲[1] 胡江[2] 李凌松[2] 

机构地区：[1]北京大学第三医院内分泌科,北京市100083 [2]北京大学干细胞研究中心,北京市100083

出　　处：《世界华人消化杂志》2005年第15期1824-1827,共4页World Chinese Journal of Digestology

基　　金：国家自然科学基金项目;No.30170443211工程建设项目人体干细胞工程学学科群资助项目;No.108-2~~

摘　　要：目的:探索一种新的人胰腺干细胞分离纯化体系,并对经这一体系所获细胞的特性进行研究.方法:首先从胎儿胰腺中分离得到胰岛样细胞簇(ICCs),进而从ICCs中得到单层上皮样细胞,并传代培养,第6代细胞通过荧光激活细胞分选(FACS)技术分选出sidepopulation(SP)细胞和非SP细胞后进行单克隆细胞培养.应用逆转录-聚合酶链反应(RT-PCR)和FACS分析,研究单克隆SP细胞的体外增殖特性和表面标志.结果:SP细胞占全部细胞的0.22%。SP细胞的克隆形成率为2.7%,而非SP细胞没有克隆形成.SP克隆能够在体外传15代以上.RT-PCR显示单克隆SP细胞可表达ATP结合盒转运子G2(ABCG2)和转录因子OCT4.FACS分析显示CD44,CDl47均为阳性,而CD34,CD38,CD45,CD71,CD133及HLA-DR均为阴性.结论:胎儿胰腺来源的单克隆SP细胞在体外具有高度的增殖潜能,并具有与骨髓间充质干细胞相同的表型特征.AIM： To find out a new method for isolation and purification of human pancreatic stem cells, and to investigate the characterization of the cells acquired by this method. METHODS： Islet-like cell clusters （ICCs） were isolated from human fetal pancreas. Monolayer epithelium-like cells were obtained from the ICCs and passaged thereafter. The side population （SP） cells or non-SP cells were sorted from these cells at the sixth generation by fluorescenceactivated cell sorting （FACS）. The culture of single SP cell or non-SP cell was assayed for clone formation. The markers of SP cells were assayed using reverse transcription polymerase chain reaction （RT-PCR） and FACS. RESULTS： The SP cells accounted for 0.22% of the total cells. The rate of clone formation was about 2.7% for the SP cells, whereas there was no clone formation for the non-SP cells. The SP cell clones were further expanded for more than 15 generations. RT-PCR analysis showed that the mRNAs of ATP-binding cassette transporter G2 （ABCG2） and transcription factor OCT4 were expressed in the cloned SP cells. FACS analysis revealed that CD44 and CD147 were positive, whereas CD34, CD38, CD45, CD71, CD133 and HLA-DR were negative. CONCLUSION： The SP cells isolated from human fetal pancreas have highly proliferative potential and share many phenotypic markers with mesenchymal stem cells derived from bone marrow.

关 键 词：单克隆SP细胞 胰腺干细胞 克隆 荧光激活细胞分选 骨髓间充质干细胞 胎儿胰腺 P细胞 表型特征 单克隆 逆转录一聚合酶链反应 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R322.11[医药卫生—基础医学]