中国组织工程研究

中国组织工程研究 ›› 2014, Vol. 18 ›› Issue (45): 7227-7232.doi: 10.3969/j.issn.2095-4344.2014.45.003

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

三维培养小鼠骨髓间充质干细胞的生物学特性

贺 鑫，李 雪，鲍会静，王仁峰，刘运德，宋士伟   

  1. 天津医科大学医学检验学院，天津市 300000
  • 出版日期:2014-11-05 发布日期:2014-11-05
  • 通讯作者: 刘运德，教授，研究生导师，天津医科大学医学检验学院，天津市 300000
  • 作者简介:贺鑫，男，1988年生，山东省淄博市人，汉族，主要从事间充质干细胞诱导耐受性树突状细胞形成机制的研究。
  • 基金资助:
    天津医科大学科学基金(2012KYQ12)，课题名称：《间充质干细胞对移植排斥反应个体树突状细胞功能影响及作用机制》；国家高技术研究发展计划项目(2012AA021003)，课题名称：《骨髓间充质干细胞诱导耐受性树突状细胞产生的策略与机制》

Effects of three-dimensional spheroid culture system on biological characteristics of mouse bone marrow mesenchymal stem cells

He Xin, Li Xue, Bao Hui-jing, Wang Ren-feng, Liu Yun-de, Song Shi-wei   

  1. School of Medical Examination, Tianjin Medical University, Tianjin 300000, China
  • Online:2014-11-05 Published:2014-11-05
  • Contact: Liu Yun-de, Professor, Master’s supervisor, School of Medical Examination, Tianjin Medical University, Tianjin 300000, China
  • About author:He Xin, School of Medical Examination, Tianjin Medical University, Tianjin 300000, China
  • Supported by:

    the Science Foundation of Tianjin Medical University, No. 2012KYQ12; the National High Technology Research and Development Program of China, No. 2012AA021003

PDF

529

被引次数

4

摘要：

背景：骨髓间充质干细胞具有自我更新和多向分化潜能而被用于多种疾病治疗及研究，但是在目前的培养条件下，其极易发生老化和自分化而失去应用价值。

 

目的：研究三维球体培养对骨髓间充质干细胞干性及衰老的影响。

 

方法：取3周龄C57/B6小鼠，分离培养骨髓间充质干细胞，分别接种于包被有壳聚糖的培养板和普通培养板。培养5 d后分别收集两组细胞，流式细胞术检测细胞表型及干性相关标志分子，应用PI和Annexin-V染色检测细胞凋亡，β-Gal染色鉴定衰老。

 

结果与结论：小鼠骨髓间充质干细胞培养第3天就可以聚集成球状生长，球体状生长的细胞干性相关标志分子CD44和Sca-1表达高于普通培养细胞，PI和Annexin-V染色可见球体培养细胞凋亡明显减少，且通过β-Gal染色可见球体状态生长的细胞无明显衰老。上述结果表明，三维球体培养技术有益于维持间充质干细胞的干性，减少其凋亡并延缓衰老。

 

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


全文链接：

关键词: 干细胞, 骨髓干细胞, 骨髓间充质干细胞, 壳聚糖, 三维球体, 干性, 凋亡, 衰老

Abstract:

BACKGROUND: Bone marrow mesenchymal stem cells have attracted widespread attention for the capabilities of self-renewal and muti-differentiation, which have been used in treatment of various diseases.

 

OBJECTIVE: To study the effect of three-dimensional spheroid culture system on the stemness and senescence of bone marrow mesenchymal stem cells.

 

METHODS: Mesechyaml stem cells were isolated from the bone marrow of C57/B6 mice, 3 weeks old, and cultured onto the culture plates coated with or without chitosan. After 5 days of culture, the cell phenotype and expression of stemness related markers CD44 and Sca-1 were analyzed by flow cytometry. PI and Annexin-V staining were used to detect cell apoptosis. Also, β-Gal staining was applied for identification of aging.

 

RESULTS AND CONCLUSION: The mouse mesenchymal stem cells began to form spheroids on day 3. The stemness-related markers, including CD44 and Sca-1, expressed higher in spheroid mesenchymal stem cells than the cells under normal culturing. Compared with the normal culture group, the apoptosis and senescence of cells from spheroid culture were lower. The results indicate that the formation of spheroids on chitosan films can increase the stemness, decrease the apoptosis and slow the senescence of mesenchymal stem cells.

 


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


全文链接：

Key words: bone marrow, mesenchymal stem cells, chitosan, apoptosis, cell aging

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