基础医学与临床 ›› 2007, Vol. 27 ›› Issue (12): 1385-1387.

• 技术与方法 • 上一篇    下一篇

促血小板生成素对骨髓c-kit+Lin-细胞体外扩增的影响

张守华 廖彩仙 张春兴 苏俊 赖勇强 周杰   

  1. 广东省南方医科大学附属南方医院肝胆外科 广东省南方医科大学附属南方医院肝胆外科 广东省南方医科大学附属南方医院肝胆外科 广东省南方医科大学附属南方医院肝胆外科 广东省南方医科大学附属南方医院肝胆外科 广东省南方医科大学附属南方医院肝胆外科
  • 收稿日期:2007-01-22 修回日期:2007-06-12 出版日期:2007-12-25 发布日期:2007-12-25
  • 通讯作者: 廖彩仙

Effects of thrombopoietin on the expansion of c-kit+Lin- cells from bone marrow in vitro

Shou-hua ZHANG, Cai-xian LIAO, Chun-xing ZHANG, Jun SU, Yong-qiang LAI, Jie ZHOU   

  1. Dept. of Hepatobiliary,Nanfang Hospital,Nanfang Medical University
  • Received:2007-01-22 Revised:2007-06-12 Online:2007-12-25 Published:2007-12-25
  • Contact: Cai-xian LIAO,

摘要: 目的 探讨在无血清条件下,血小板生成素(TPO)对c-kit+Lin-细胞增殖的影响。方法 经免疫磁珠分离小鼠骨髓c-kit+Lin-细胞,研究在干细胞因子(SCF)、FLt-3配基(FL)和白血病抑制因子(LIF)组合中添加不同浓度的TPO,7d后检测细胞总数,c-kit+Lin-细胞扩增倍数,并采用 Annexin-V检测各组细胞凋亡率。结果 各组均能显著扩增c-kit+Lin-细胞,扩增倍数为4~7倍不等。当SCF+FL+LIF中加入 50μg/L TPO时,c-kit+Lin-细胞及细胞总数扩增为7.07倍和40.19倍,比未加时明显增多,而凋亡率无明显差异。但随剂量加大,c-kit+Lin-细胞扩增下降,细胞凋亡率上升,200μg/L TPO组凋亡率为33.49%,c-kit+Lin-细胞扩增倍数仅为4.11倍。结论 TPO可协同SCF+FL+LIF使c-kit+Lin-细胞增殖,但过高剂量会诱导其分化和凋亡,以50μg/L浓度扩增效果最佳。

关键词: 血小板生成素, 骨髓c-kit+Lin-细胞, 肝干细胞, 扩增

Abstract: Objective To elucidate the effect of thrombopoietin (TPO) in vitro expansion of c-kit+Lin- cells from bone marrow . Methods c-kit+Lin- cells were isolated by using a high-gradient magnetic cell sorting system (MACS) , and expanded with combinations of stem cell factor(SCF), FLt-3 ligand (FL) ,leukemia inhibitor factor(LIF)and different concentration of thrombopoietin (TPO) for 7days in a liquid culture system. Totall cells and Annexin-V positive cells were studied by fluorescence-activated cell sorter ( FACS) . Results Each group could expand c-kit+Lin- cells effectively and rapidly,and the numbers of cells expanded were 4~7 times compared with the beginning's number. Added 50μg/L TPO into SCF+FL+LIF increased expasions of c-kit+Lin- and total cells by 7.07 and 40.19 times,which was more than the group that had no TPO。But when the concentration of TPO increased,the c-kit+Lin- cells decreased,and the apoptosis rate rised. Conclusion It showed TPO synergistic actions with SCF+FL+LIF to expand c-kit+Lin- cells but too much TPO could induce it differentiation and apoptosis.50μg/L TPO was the best concentration to expand c-kit+Lin- cells.

Key words: Thrombopoietin, c-kit+Lin-cells in bone marrow, hepatic stem cell, expand