国际口腔医学杂志 ›› 2010, Vol. 37 ›› Issue (3): 272-272~274.doi: 10.3969/j.issn.1673-5749.2010.03.007

• 论著 • 上一篇    下一篇

骨髓基质干细胞的体外培养及其细胞片层制备

荆恒,李宁毅,谭帅,高振华,陈立强   

  1. 青岛大学医学院附属医院口腔颌面外科 山东 青岛 266003
  • 收稿日期:2009-12-22 修回日期:2010-03-11 出版日期:2010-05-20 发布日期:2010-05-20
  • 通讯作者: 李宁毅

Culture in vitro and the fabrication of cell sheet for bone marrow stem cells

JING Heng, LI Ning-yi, TAN Shuai, GAO Zhen-hua, CHEN Li-Qiang   

  1. Dept. of Oral and Maxillofacial Surgery, Affiliated Hospital, Medical College, Qi
  • Received:2009-12-22 Revised:2010-03-11 Online:2010-05-20 Published:2010-05-20
  • Contact: LI Ning-yi

摘要: 目的 探讨犬骨髓基质干细胞(BMSCs)的体外培养及成骨向诱导后细胞片层的制备方法。方法 密度梯度离心法分离培养犬BMSCs,向成骨细胞方向诱导分化后,倒置相差显微镜和扫描电镜下进行形态学观察;将诱导后的BMSCs 接种至温度反应性培养皿中,37 ℃、5%CO2 饱和湿度培养,然后降温至20 ℃制备BMSCs 的细胞片层。结果 原代培养的BMSCs 呈长梭形,漩涡状生长;向成骨细胞诱导后的BMSCs 呈卵圆形或多角形,当温度降至20 ℃时,BMSCs 从温度反应性培养皿上完全脱落。扫描电镜下观察可见到富含细胞外基质的完整BMSCs 细胞片层。结论 应用密度梯度离心法可有效获取BMSCs,并向成骨细胞诱导分化;应用温度反应性培养皿能获取完整的BMSCs 细胞片层可包裹支架,并用于功能性成骨。

关键词: 骨髓基质干细胞, 细胞片层, 温度反应性培养皿

Abstract: Objective To explore the methods of culture and induction of bone marrow stem cells(BMSCs) in vitro and fabrication of cell sheet of BMSCs. Methods Bone marrow stem cells were isolated with the method of density gradient centrifugation from canine and cultured. BMSCs were induced to differentiate to osteoblasts. The BMSCs were observed under inverted microscope and scanning electron microscope. The BMSCs after induced were transferred to the temperature-responsive culture dishes and cultured in incubator at 37 ℃, 5% CO2 and saturated humidity. The temperature of the incubator was changed from 37 ℃ to 20 ℃, to fabricate the cell sheet of BMSCs. Results BMSCs was fusiform and grew in whirl-like arrangement while the BMSCs induced were oval or polygonal; when the temperature dropped to 20 ℃, bone marrow stem cell sheet detached automatically from the temperature-responsive culture dishes. Conclusion BMSCs can be effectively obtained with the method of density
gradient centrifugation and induced to differentiate into osteoblasts. Bone marrow stem cell sheet layer can be used to wrap the scaffold for construction of functional osteogenesis.

Key words: bone marrow stem cells, cell sheet, temperature-responsive culture dishes

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[1] 程谷综述 李祖兵审校. 骨组织工程中的检测方法及其效果评估展望[J]. 国际口腔医学杂志, 2010, 37(5): 593-596.
[2] 荆恒, 陈欣综述 李宁毅审校. 细胞片层技术的研究进展[J]. 国际口腔医学杂志, 2010, 37(01): 71-71~73.
[3] 蒋欣泉,陈传俊. 应用骨髓基质干细胞修复骨缺损的研究进展[J]. 国际口腔医学杂志, 2002, 29(01): -.
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