模拟微重力条件下对WB-F344肝干细胞表征分子表达的影响

研究大鼠WB-F344肝干细胞在旋转式细胞培养系统（RCCS）中培养进行细胞大规模扩增并保持干细胞的特性的可能性，为干细胞治疗疾病及肝组织工程提供理想的细胞来源。以WB-F344肝干细胞在RCCS中培养,以平面单层培养为对照，在培养后不同时间分别进行形态观察、流式细胞仪测细胞周期、逆转录-聚合酶链反应（RT-PCR）检测肝干细胞特异性基因甲胎蛋白（AFP）和白蛋白（ALB）的表达, 免疫荧光染色检测 AFP、ALB蛋白的表达。结果表明，RCCS培养的WB-F344细胞粘附在Cytodex-3微载体上状态生长良好，细胞增殖较平面培养有明显增加；RT-PCR和免疫荧光染色检测结果一致：模拟微重力培养组AFP的mRNA表达强度及AFP阳性细胞均显著高于平面培养组，而ALB mRNA表达强度和ALB阳性细胞均低于对照组。说明模拟微重力 培养条件下，能较好的维持肝干细胞特性，进一步证明我们建立的这种培养体系是成功的，是一种理想的肝干细胞培养模式。

The Affection of Molecular Expression in Three-dimensional Cultivating WB-F344 Cells

To study hepatic stem cells(HSCs) proliferation and characteristic in rotary cell culture system(RCCS) for ideal cell origin of liver tissue engineering and clinic liver diseases stem cells therapy.WB-F344,a kind of rat liver stem cell,was cultured in RCCS to investigate the effect of RCCS onhepatic stem cell characteristics,and the condition of two-dimensional(2D) monolayer cultivating as control.To analyze the changing ofhSCs characteristics by morphological observation,flow cytometry(FACS),RT-PCR and immunofluorescence.To the results,demonstrated that cells were grow well and packed together densely to form irregular aggregates in RCCS,and proliferation of thehepatic stem cells was superior with control,mRNA of AFP increased while ALB decreased,the protein of AFP expression increased while ALB decreased comparing to control.Indicating RCCS could afford appropriate environment to sustain the stem cells character of WB-F344 cells.