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小鼠羊水间充质干细胞的分离、培养及鉴定

吴航飞 王康淳 潘崎 程颖

吴航飞, 王康淳, 潘崎, 等. 小鼠羊水间充质干细胞的分离、培养及鉴定[J]. 器官移植, 2022, 13(1): 67-73. doi: 10.3969/j.issn.1674-7445.2022.01.011
引用本文: 吴航飞, 王康淳, 潘崎, 等. 小鼠羊水间充质干细胞的分离、培养及鉴定[J]. 器官移植, 2022, 13(1): 67-73. doi: 10.3969/j.issn.1674-7445.2022.01.011
Wu Hangfei, Wang Kangchun, Pan Qi, et al. Isolation, culture and identification of mouse amniotic fluid-derived mesenchymal stem cells[J]. ORGAN TRANSPLANTATION, 2022, 13(1): 67-73. doi: 10.3969/j.issn.1674-7445.2022.01.011
Citation: Wu Hangfei, Wang Kangchun, Pan Qi, et al. Isolation, culture and identification of mouse amniotic fluid-derived mesenchymal stem cells[J]. ORGAN TRANSPLANTATION, 2022, 13(1): 67-73. doi: 10.3969/j.issn.1674-7445.2022.01.011

小鼠羊水间充质干细胞的分离、培养及鉴定

doi: 10.3969/j.issn.1674-7445.2022.01.011
基金项目: 

辽宁省教育厅科学研究项目 FWZR2020002

详细信息
    作者简介:

    吴航飞,男,1996年生,硕士,研究方向为器官移植,Email:wu452678240@163.com

    通讯作者:

    程颖,女,1975年生,博士,研究员,研究方向为器官移植,Email:chengying75@sina.com

  • 中图分类号: R617, R392.2

Isolation, culture and identification of mouse amniotic fluid-derived mesenchymal stem cells

More Information
  • 摘要:   目的  探讨小鼠羊水间充质干细胞(AF-MSC)的分离、培养及鉴定。  方法  在无菌条件下获取孕鼠子宫,收集羊水后进行过滤和离心,对沉淀细胞团进行培养并传代。观察AF-MSC的形态,分析AF-MSC的增殖特点,采用流式细胞术鉴定AF-MSC的表面标志物,检测AF-MSC的三系分化能力及冷冻复苏后的细胞活力。  结果  小鼠AF-MSC呈典型的梭形,融合度 > 80%时会出现典型的漩涡状结构。小鼠AF-MSC传代培养无明显潜伏期,培养2~3 d进入对数增长期,增长速度最快,之后增殖速度减慢,进入平台期。AF-MSC表达干细胞抗原(Sca)-1、CD29、CD44,不表达CD34、CD45。小鼠AF-MSC成骨分化后,矿化结晶被茜素红染成深红色的点状;成软骨分化后,分泌的酸性粘多糖被阿利新蓝染成淡蓝色;成脂分化后,胞质脂滴被油红O染成红色。细胞冷冻复苏后存活率 > 95%,生长状态良好,6 d时增殖能力高于冻存前(P < 0.05),其他时间增殖能力与冻存前比较,差异无统计学意义(均为P > 0.05)。  结论  本实验成功分离了小鼠AF-MSC,过程简便、成本低,且分离的细胞可随传代次数的增加而纯化,冻存不影响其增殖能力。

     

  • 图  1  AF-MSC的形态

    注:A、C、E图分别为P0、P2、P4 AF-MSC形态(×100);B、D、F图为P0、P2、P4 AF-MSC形态(×200)。

    Figure  1.  The morphology of AF-MSC

    图  2  AF-MSC的增殖曲线

    注:与P3比较,aP<0.05/3。

    Figure  2.  Proliferation curves of AF-MSC

    图  3  AF-MSC表面标志物的流式细胞图

    Figure  3.  Flow cytometry of surface markers of AF-MSC

    图  4  AF-MSC的三系分化染色图

    注:A图为AF-MSC茜素红染色图(×200);B图为AF-MSC阿利新蓝染色图(×100);C图为AF-MSC油红O染色图(×200)。

    Figure  4.  The trilineage differentiation figures of AF-MSC

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出版历程
  • 收稿日期:  2021-09-22
  • 网络出版日期:  2022-01-12
  • 刊出日期:  2022-01-15

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