中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (45): 8361-8365.doi: 10.3969/j.issn.1673-8225.2011.45.001

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

流动优化骨髓间充质干细胞的分化状况

刘  琳1，杨思远2，胡选义2，赁  可3，石应康3   

  1. 1贵州省人民医院呼吸内科，贵州省贵阳市 550002
    2贵阳医学院附属医院心脏外科，贵州省贵阳市550004
    3四川大学华西医院胸心血管外科，四川省成都市  610041
  • 收稿日期:2011-04-14 修回日期:2011-05-26 出版日期:2011-11-05 发布日期:2011-11-05
  • 通讯作者: 杨思远，博士，副主任医师，贵阳医学院附属医院心脏外科，贵州省贵阳市550004 wo418@tom.com
  • 作者简介:刘琳★，女，1979年生，四川省自贡市人，汉族，2008年遵义医学院呼吸内科学毕业，硕士，主治医师，主要从事心肺慢性疾病基础及临床方面研究。
  • 基金资助:

    国家自然科学基金(10402026)；贵州省优秀科技教育人才省长专项资金(黔省专合字200891号)；贵阳医学院附属医院博士基金。

Multipotent differentiation status of rat bone marrow mesenchymal stem cells in vitro after shear stress loading

Liu Lin1, Yang Si-yuan2, Hu Xuan-yi2, Lin Ke3, Shi Ying-kang3   

  1. 1Department of Respiratory Medicine, Guizhou Provincial People’s Hospital, Guiyang  50002, Guizhou Province, China
    2Department of Cardiac Surgery, the Affiliated Hospital of Guiyang Medical College, Guiyang  550004, Guizhou Province, China
    3Department of Thoracic-Cardiac Surgery, West China Hospital, Sichuan University, Chengdu  610041, Sichuan Province, China
  • Received:2011-04-14 Revised:2011-05-26 Online:2011-11-05 Published:2011-11-05
  • Contact: Yang Si-yuan, Doctor, Associate chief physician, Department of Cardiac Surgery, the Affiliated Hospital of Guiyang Medical College, Guiyang 550004, Guizhou Province, China wo418@tom.com
  • About author:Liu Lin★, Master, Attending physician, Department of Respiratory Medicine, Guizhou Provincial People’s Hospital, Guiyang 50002, Guizhou Province, China
  • Supported by:

    the National Natural Science Foundation of China, No. 10402026*; Guizhou Provincial Governor Special Fund for Scientific and Educational Talents, No. 200891*; Doctor Foundation of the Affiliated Hospital of Guiyang Medical College*

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摘要：

背景：间充质干细胞在成体动物骨髓中含量极低。
目的：观察成年大鼠骨髓间充质干细胞经生理范围流动切应力作用后，其体外生长规律、增殖及定向分化为成骨细胞及脂肪细胞的能力。
方法：在体外取得SD大鼠骨髓原代间充质干细胞，利用平板流室系统加载1 Pa切应力，对其进行培养及扩增，并分别向成骨细胞及脂肪细胞定向诱导。
结果与结论：原代间充质干细胞为长梭形或多角形，集落样生长；经流动加载并传代后细胞生长速度加快，不以集落方式生长，而是均匀分布。细胞体积明显增大，多数为长梭形或多角形；生长曲线显示各代细胞有类似的生长规律；激光共聚焦显微镜显示CD44和CD90阳性，CD31及CD45阴性；向成骨细胞及脂肪细胞诱导14 d后，Von Kossa染色及油红O染色均为阳性，流动优化后骨髓间充质干细胞保持了体外大量增殖及多向分化潜能。

关键词: 流动切应力, 骨髓, 大鼠Spragua-Dawley, 间充质干细胞, 诱导分化

Abstract:

BACKGROUND: The number of mesenchymal stem cells in the bone marrow of adult animals is little.
OBJECTIVE: To investigate whether shear stress could modify the abilities of growth, amplification and multi-potent differentiation of bone marrow mesenchymal stem cells or not.
METHODS: SD rat bone marrow mesenchymal stem cells were isolated, cultured and amplified in vitro by Percoll density gradient centrifugation and adherent culture. 1 Pa shear stress was loaded on primary passage mesenchymal stem cells with a flow chamber system for osteogenic and adipogenic differentiation.
RESULTS AND CONCLUSION: The primary cells were mostly spindle-like in shape, like the fibroblast cells, and formatted clones. After the cells were loaded with shear stress and then passaged, they grew faster and bigger, and had the same morphologic characters similar as primary mesenchymal stem cells. The growth curves showed the growth regularity of all passage cells was similar, the laser scanning confocal microscope showed the CD41 and CD90 were positive, CD31 and CD45 were negative. After the cells were induced for 14 days, the results of Von Kossa strain and oil red O strain were both positive. The abilities of growth, amplification and multi-potent differentiation of bone marrow mesenchymal stem cells maintained after shear stress loading.

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