小鼠胎肝细胞透明质酸3D培养体系的建立
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1.广西医科大学,南宁 530021;2.北京军事科学院 军事医学研究院 卫生勤务与血液研究所 再生医学研究室,北京 100039

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Stem-cell Derived, Functional Mouse Hepatic Organoids In Hyaluronan Hydrogels
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1.Guangxi Medical University, Nanning 530021, China;2.Regenerative Medicine Lab, Institute of Health Service and Transfusion Medicine, Beijing 100039, China

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    摘要:

    目的 利用透明质酸建立小鼠胎肝细胞3D培养体系。 方法 分离获得胚胎12-14天胎肝细胞,利用KM培养基进行初步2D肝干/祖细胞的筛选培养,并利用透明质酸及KM培养基配制水凝胶建立3D细胞培养体系。 结果 胎肝细胞在2D体系中呈现克隆状生长。分离培养获得的肝干/祖细胞克隆在透明质酸建立的3D培养体系保持增殖活性,并进一步获得肝细胞功能特性,表现为3D培养上清中白蛋白合成和尿素水平显著增加。Q-PCR结果显示随着3D培养时间的延长,其肝细胞干性标志如AFP、CK19、EpCAM、Prox1等表达水平都大幅度降低且接近成年小鼠肝脏表达水平。 结论 本研究成功建立基于透明质酸的小鼠胎肝细胞的3D无血清培养体系,并可促进小鼠胎肝细胞肝细胞功能进一步成熟。

    Abstract:

    Objective: To establish a 3D organoid culture system of mouse fetal liver cell using hyaluronic acid. Method: The fetal liver cells isolated from 12 to 14 days mouse embryos were preliminary screening of 2D liver stem/progenitor cells by Kubota’s Medium (KM) and 3D organoid culture in hyaluronan hydrogel. Result: Fetal liver cells grew as clones in 2D cell culture system. Liver stem/progenitor cell colonies maintained proliferation activity. The albumin and urea levels in supernatant increased significantly, which indicated hepatic functional maturation of the cell colonies in 3D hyaluronic hydrogels. The results of Q-PCR showed that the expression levels of liver stem/progenitor markers, such as AFP, CK19, EpCAM and Prox1, significantly reduced and were close to the expression level in adult mouse liver. Conclusion: In this study, stem-cell derived, functional mouse hepatic organoids in hyaluronan hydrogels was successfully established.

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颜洁心,王怡,王韫芳.小鼠胎肝细胞透明质酸3D培养体系的建立[J].生物化学与生物物理进展,2019,46(9):27-34

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历史
  • 收稿日期:2018-03-08
  • 最后修改日期:2018-04-09
  • 接受日期:2018-04-11
  • 在线发布日期: 2018-05-18
  • 出版日期: 2019-09-20