小鼠肺间充质干细胞的分离、培养和鉴定
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中国医学科学院医学实验动物研究所,北京协和医学院比较医学中心,国家卫生健康委员会人类疾病比较医学重点实验室,国家中医药管理局人类疾病动物模型三级实验室,北京 100021

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R-33

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Isolation, culture, and characterization of mouse lung-resident mesenchymal stem cells
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Institute of Laboratory Key Laboratory of Human Disease Sciences, Chinese Academy of Medical Sciences(CAMS); Comparative Medicine Center, Peking Union Medical College(PUMC), Key Laboratory of Human Disease Comparative Medicine, National Health Commission of the People’s Republic of China; Key Laboratory of Human Disease Key Laboratory of Human Disease Models, State Administration of Traditional Chineses Medicine, Beijing 100021, China

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    摘要:

    目的 建立稳定的小鼠肺间充质干细胞分离、培养、鉴定技术,利用小鼠肺间充质干细胞研究肺组织修复和再生机制。 方法 无菌分离小鼠肺,采用胶原酶消化和组织贴壁法分离培养肺间充质干细胞,观察细胞形态和生长特性,流式分析细胞表面标记,进行体外成脂、成骨诱导分化。 结果 两种方法都成功分离培养了肺间充质干细胞,流式分析显示 Sca-1、CD90、CD29、CDCD44 均为高表达,CD31、CD45 均为低表达。体外诱导后,肺间充质干细胞可以分化成脂肪细胞和成骨细胞。 结论 本研究利用两种方法分离培养小鼠肺间充质干细胞,两种分离方法获得的细胞均具有间充质干细胞类似的表面标记物,具有成脂、成骨分化能力。

    Abstract:

    Objective To establish a stable method for the isolation, culture, and characterization of mouse lung- resident mesenchymal stem cells, which will then be used to study lung repair and regeneration mechanisms. Methods Mouse lungs were obtained via germ-free procedures and were mechanically minced and / or digested with collagenase. Cell morphology was observed by phase-contrast microscopy, cell surface markers were detected by flow cytometry, and cells were induced to differentiate into adipocytes or osteoblasts. Results We successfully isolated and cultured mouse lung- resident mesenchymal stem cells. FACS analysis demonstrated high expressions of Sca-1, CD90, CD29, and CD44, and low expressions of CD31 and CD45. The lung-resident stem cells differentiated into adipocytes or osteoblasts. Conclusions We isolated cells that expressed mesenchymal stem cell markers and had multi-directional differentiation ability.

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石桂英,黄艺滢,雷雪裴,路亚岚,白 琳.小鼠肺间充质干细胞的分离、培养和鉴定[J].中国比较医学杂志,2021,31(8):16~20.

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  • 收稿日期:2020-11-05
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  • 在线发布日期: 2021-09-26
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