Chinese Journal of Tissue Engineering Research ›› 2012, Vol. 16 ›› Issue (1): 56-60.doi: 10.3969/j.issn.1673-8225.2012.01.012

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Effects of human embryonic lung fibroblasts feeder layer transfected with leukemia inhibitory factor gene on the proliferation of umbilical cord blood CD34+ cells in vitro

Xia Wei1, Yu Xin1, Jiang Jian1, Tian Li-na2, Chen Yao2, Miao Jing-cheng2   

  1. 1Key Laboratory of Blood Transfusion, Wuxi Red Cross Blood Centre, Wuxi  214021, Jiangsu Province, China; 2Department of Cell and Molecular Biology, Medicine School of Soochow University, Suzhou  215123, Jiangsu Province, China
  • Received:2011-05-13 Revised:2011-07-15 Online:2012-01-01 Published:2012-01-01
  • Contact: Yu Xin, Master, Associate chief technician, Key Laboratory of Blood Transfusion, Wuxi Red Cross Blood Centre, Wuxi 214021, Jiangsu Province, China yu_heart@sina.com
  • About author:Xia Wei, Associate chief technician, Key Laboratory of Blood Transfusion, Wuxi Red Cross Blood Centre, Wuxi 214021, Jiangsu Province, China
  • Supported by:

    the Research Foundation of Jiangsu Province Health Department, No. H200955*; the Science and Technology Social Development Program of Wuxi City, No. CSZ00962*

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Abstract:

BACKGROUND: The number of hematopoietic cells in single copy cord blood is limited, and it is difficult to meet the needs of adults. How to effectively amplify cord blood hematopoietic stem/progenitor cells (HSPCs) is a hot research issue currently.
OBJECTIVE: To construct gene-modified human embryonic lung fibroblast cells using human leukemia inhibitory factor (hLIF) and to observe the effect of transgenic cells on the amplification of HSPCs in vitro.
METHODS: The feeder layer trancfected with recombinat adenouirus Ad-hLIF was established and objective gene was detected by using RT-PCR and ELISA. Umbilical cord blood CD34+ HSPC were separated using magnetic-activated cell sorting and the purity was detected using flow cytometry. Homing ability of amplified HSPC in vitro was detected by transmembrane migration assay.
RESULTS AND CONCLUSION: Transgenic feeder cells were constructed successfully, and the expression of objective gene was verified by RT-PCR and ELISA. After culturing with feeder layer cells for seven days, the number of CD34+ HSPCs in the group containing hLIF was 15.73 much more than that in the group without hLIF. The expression rate of surface adhesion molecules was also still great. In vitro migration assay showed that the induced migration rate of hematopoietic cells in co-culturing with transgenic feeder layer was significantly higher than that of the control group, with a better homing ability. The feeder layer cells of adenovirus vector transfected with hLIF gene amplify umbilical cord blood CD34+ HSPC effectively in vitro and delay differentiation; what’s more, the CD34+ cells retains a great homing ability after amplifying in vitro.

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