基础医学与临床 ›› 2009, Vol. 29 ›› Issue (9): 911-915.

• 研究论文 • 上一篇    下一篇

转基因饲养层细胞体外促进脐带血CD34+HSPC增殖研究

胡志清 杨吉成 盛伟华 井莹莹 苗莉 张日 朱南康 缪竞诚   

  1. 苏州大学医学部
  • 收稿日期:2008-10-06 修回日期:2008-11-22 出版日期:2009-09-20 发布日期:2009-09-20

Study on the function of Transgenic cell line as a feeder layer on UCB CD34+ HSPC ex vivo amplification

Zhi-qing HU, Ji-cheng YANG, Wei-hua SHENG, Ying-ying JING, Li MIAO, Ri ZHANG, Nan-kang ZHU, Jing-cheng MIAO   

  1. Medical College, Soochow University
  • Received:2008-10-06 Revised:2008-11-22 Online:2009-09-20 Published:2009-09-20

摘要: 目的 构建转hOSM基因细胞系作为饲养层细胞,观察其对脐带血CD34+HSPC的扩增作用。方法 用分子生物学技术建立转hOSM基因细胞系,免疫磁珠法分离脐带血CD34+HSPC,将其与饲养层细胞共培养7d后,流式细胞术检测其增殖,将扩增后染色的CD34+细胞移植入SCID小鼠体内,应用荧光显微镜和RT-PCR法观察移植效果。结果 CD34+HSPC与饲养层细胞共培养后扩增了9.4倍,其表面归巢相关分子CXCR4和CD54阳性率仍较高,植入小鼠体内4周后,可成功归巢到小鼠骨髓。结论 建立的转hOSM基因饲养层细胞可以有效的扩增CD34+HSPC,并维持其较高的移植效率和造血活性。

Abstract: Objective To establish the transgenic cell line of hOSM gene and observe its effect on the proliferation of umbilical cord blood(UCB)CD34+ HSPC as a feeder layer. Methods A transgenic cell line was established by molecular biology method and CD34+ HSPCs were separated by magnetic-activated cell sorting. After 7 days' co-culture, the amplification effect was evaluated by flow cytometry. Then CD34+ cells stained by CFDA SE were injected into the SCID mice with sublethally irradiation and the effect of transplation was observed by fluorescence microscopy and RT-PCR. Results The number of CD34+ cells increased by 9.6 times and the expression rate of homing-ralated molecules was still higher than 50%. Four weeks after transplantion, CD34+ cells were found in bone marrow of SCID mice. Conclusion The transgenic cell line of hOSM gene as a feeder layer can effectively amplify UCB CD34+ HSPC ex vivo and suppress its differentiation. Meanwhile, they have high transplantion efficacy and haematogenesis activity.