《中国康复理论与实践》

《中国康复理论与实践》 ›› 2008, Vol. 14 ›› Issue (01): 21-23.

• 基础研究 • 上一篇    下一篇

兔干细胞的体外培养和碱性成纤维细胞生长因子对其代谢的影响

马舟涌; 李放; 余兆仲; 任大江; 陈晓斌   

  1. 北京军区总医院骨科,北京市100700
  • 收稿日期:2007-07-09 出版日期:2008-01-01 发布日期:2008-01-01

Culturing Rabbit Stem Cells in Vitro and Its Anabolism under Basic Fibroblast Growth Factor

MA Zhou-yong, LI Fang, YU Zhao-zhong, et al   

  1. the Forth Military Medical University, Beijing Army General Hospital,Beijing 100700, China
  • Received:2007-07-09 Published:2008-01-01 Online:2008-01-01

PDF (PC)

被引次数

10

摘要： 目的探索体外培养的骨髓间充质干细胞(BMSCs)和脂肪干细胞(ASCs)变化,以及碱性成纤维细胞生长因子(bF-GF)对其代谢的影响。方法分别用DMEM、DMEM/F12(2∶1)、α-MEM培养兔BMSCs和ASCs。将体外培养的第3代兔BMSCs和ASCs各分为2组后培养到第3代。A组:软骨诱导培养基(CM)+ASCs,B组:CM+ASCs+bFGF5ng/ml,C组:BMSCs+CM,D组:BMSCs+CM+bFGF5ng/ml。倒置显微镜观察细胞形态学变化,测定35SO42-摄入量及羟脯氨酸浓度。结果α-MEM培养基比DMEM、DMEM/F12(2∶1)制备的BMSCs和ASCs所需时间缩短。单层培养的BMSCs和ASCs增殖较快,贴壁较牢且生成细胞集落,但经CM诱导后,细胞增殖较慢,贴壁不牢,且细胞没有集落生成。诱导后的BMSCs和ASCs的形状趋向于类圆形,B组和D的细胞数量比A组和C组的细胞多,B组、D组分别比A组、C组表达羟脯氨酸增加、35SO42-摄入量增加,而D组羟脯氨酸表达和35SO42-摄入量与B组无显著性差异。结论添加了bFGF的CM诱导液诱导的兔干细胞生长较好,代谢增加。

关键词: 髓核, 细胞培养, 脂肪干细胞(ASCs), 骨髓间充质干细胞(BMSCs), 组织工程

Abstract: Objective To explore the changes of rabbit adipose stem cells(ASCs)and bone mesenchymal stem cells(BMSCs)cultured in vitro and the anabolism under basic fibroblast growth factor(bFGF).Methods BMSCs and ASCs were cultured with DMEM,DMEM/F12(2∶1)or α-MEM respectively.The 3rd generation ASCs and BMSCs were divided into 2 groups respectively:group A:ASCs cultured in chondrogenic medium(CM),group B:ASCs cultured in CM supplemented with bFGF 5 ng/ml,group C:BMSCs cultured in CM,group D:BMSCs cultured in CM supplemented with bFGF 5 ng/ml.Morphological changes were observed under inverted microscope.The 35SO42-incorporation and total hydroxyproline were measured.Results BMSCs and ASCs showed much higher growth rate when cultured in α-MEM medium comparison with that in DMEM or in DMEM/F12(2:1).Both stem cells attachment cultured in monolayer greatly increased and cell clones were abundant,while the cells attachment became rather difficult and cell clones were less after cutured in CM.All stem cells possessed a round-like morphology,and the cells in group B and D were more than that in the other 2 groups.The 35SO42-incorporation and total hydroxyproline synthesis of group B or D increased compared with that of group A or C,but there was no diference between group D and B.Conclusion The rabbit ASCs and BMSCs cultured in CM suppling with bFGF grow well and their metabolism increased.

Key words: nucleus pulposus, cell culture, adipose stem cells(ASCs), bone mesenchymal stem cells(BMSCs), tissue engineering