中国组织工程研究

中国组织工程研究 ›› 2018, Vol. 22 ›› Issue (33): 5255-5258.doi: 10.3969/j.issn.2095-4344.0658

• 脐带脐血干细胞 umbilical cord blood stem cells • 上一篇    下一篇

混合人AB血清体外有效扩增脐带间充质干细胞的可行性

侯 萍，李晓丰，李伟伟，李剑平   

  1. 辽宁省血液中心，沈阳中心血站，输血医学研究所，辽宁省沈阳市 110044
  • 修回日期:2018-07-26 出版日期:2018-11-28 发布日期:2018-11-28
  • 通讯作者: 李剑平，主任医师，辽宁省血液中心，沈阳中心血站，输血医学研究所，辽宁省沈阳市 110044
  • 作者简介:侯萍，女，1977年生，辽宁省沈阳市人，汉族，2005年中国医科大学毕业，硕士，副主任医师，主要从事干细胞研究。
  • 基金资助:

    2017年辽宁省博士科研启动基金指导计划项目(21070520339)

Feasibility of mixed human AB serum to expand human umbilical cord mesenchymal stem cells in vitro

Hou Ping, Li Xiao-feng, Li Wei-wei, Li Jian-ping   

  1. Shenyang Center Blood (Liaoning Blood Center), Shenyang 110044, Liaoning Province, China
  • Revised:2018-07-26 Online:2018-11-28 Published:2018-11-28
  • Contact: Li Jian-ping, Chief physician, Shenyang Center Blood (Liaoning Blood Center), Shenyang 110044, Liaoning Province, China
  • About author:Hou Ping, Master, Associate chief physician, Shenyang Center Blood (Liaoning Blood Center), Shenyang 110044, Liaoning Province, China
  • Supported by:

    the 2017 Liaoning Provincial Doctoral Research Startup Fund Guidance Project, No. 21070520339

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摘要：

文章快速阅读：

文题释义：
人脐带间充质干细胞：具有来源丰富、多向分化潜能和免疫调节功能，是细胞治疗和组织工程领域最有潜力的候选种子细胞之一，目前多采用体积分数为10%胎牛血清或者多种细胞因子组合或者商品化的无血清间充质干细胞专用培养基培养人脐带间充质干细胞。
间充质干细胞专用培养基：是商品化的间充质干细胞专用培养基，未添加血清，添加各种生长因子，使用方便，但价格相对昂贵。

 

摘要
背景：目前人脐带间充质干细胞培养多采用体积分数为10%胎牛血清或者多种细胞因子组合或者商品化的无血清间充质干细胞专用培养基进行培养，但还没有一种价格低廉、适用临床和相对安全用于患者的标准化培养体系。
目的：探讨混合人AB血清用于培养人脐带间充质干细胞的可行性。
方法：采用组织块贴壁法分离培养人脐带间充质干细胞，分别用含体积分数为10%人AB血清的基础培养基以及无血清间充质干细胞专用培养基进行培养，检测2种培养体系中人脐带间充质干细胞的细胞形态、免疫表型、细胞周期等基础指标。
结果与结论：①2种培养体系中的人脐带间充质干细胞形态没有显著差异，均为长梭形，但混合人AB血清培养的人脐带间充质干细胞黏附力更强；②流式细胞检测结果显示，2种培养体系中人脐带间充质干细胞均高表达CD73和CD90，低表达CD45和HLA-DR，且表达量差异无显著性意义(P > 0.05)；③细胞周期结果显示，2种培养体系中多数人脐带间充质干细胞处于G0-G1期以及S期，DNA指数、变异系数均在正常范围之内；④实验证实，混合人AB血清可作为临床级别的人源血清添加到基础培养基中安全使用。

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程
ORCID: 0000-0002-8643-3282(侯萍)

关键词: 人脐带间充质干细胞, 混合人AB血清, 细胞形态, 免疫表型, 细胞周期, 干细胞

Abstract:

BACKGROUND: At present, human umbilical cord mesenchymal stem cells (HUMSCs) are mainly cultured in a medium containing 10% fetal bovine serum or a combination of various cytokines or cultured in commercial serum-free medium, but there is no low-cost, relatively safe standardized culture system applicable to the clinical utility.
OBJECTIVE: To explore the effective expansion of HUMSCs by mixed human AB serum.
METHODS: HUMSCs were isolated by tissue explant method, and then cultured in the basal medium containing 10% mixed human AB serum or in the serum-free medium. Differences between the two culture systems were then detected, involving the morphological characteristics, proliferative capacity and cell cycle.
RESULTS AND CONCLUSION: There was no significant difference in cell morphology between the two culture systems, and all the cultured cells were long shuttle-shaped. However, HUMSCs cultured with human AB serum had stronger adhesion. Flow cytometry results showed that HUMSCs highly expressed CD73 and CD90, but lowly expressed CD45 and HLA-DR, and the expression levels had no significant difference in both culture systems (P > 0.05). The cell cycle results showed that most cells were in G0-G1 and S phases, and the DNA index and coefficient of variation values in both culture systems were within the normal range. To conclude, the mixed human AB serum is considered to be a human-derived serum of clinical grade for safe use in the HUMSCs culture medium. 

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: ABO Blood-Group System, Serum, Umbilical Cord, Mesenchymal Stem Cells, Immunophenotyping, Cell Cycle, Tissue Engineering

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