Expansion of CD34+ cells from human umbilical cord blood by FL and/or TPO gene transfected human marrow stromal cell lines |
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Authors: | Yi Zhang Peixian Tang Ying Jin Xiusen Li Shuangxi Zhang Ying Wu Ning Mao |
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Institution: | (1) Institute of Basic Medical Sciences, Academy of Military Medical Sciences, 100850 Beijing, China |
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Abstract: | To elucidate the effect of gene transfected marrow stromal cell on expansion of human cord blood CD34+ cells, a culture system was established in which FL and TPO genes were transfected into human stromal cell line HFCL. To
establish gene transfected stromal cells co-culture system, cord blood CD34+ cells were purified by using a magnetic beads sorting system. The number of all cells and the number of CD34+ cells and CFC (CFU-GM and BFU-E) were counted in different culture systems. The results showed that in all 8 culture systems,
SCF+IL-3+HFT manifested the most potent combination, with the number of total nucleated cells increasing by (893.3 ±52.1)-fold,
total progenitor cells (CFC) by (74.5 ±5.2)-fold and CD34+ cells by 15.7-fold. Maximal expansions of CFC and CD34+ cells were observed at the end of the second week of culture. Within 14 days of culture, (78.1 ± 5.5)-fold and (57.0 ± 19.7)-fold
increases in CFU-GM and BFU-E were obtained. Moreover, generation of LTC-IC from amplified CD34+ cells within 28 days was found only in two combinations, i.e. SCF+IL-3+FL+TPO and SCF+IL-3+HFT, and there was no significant
difference between these two groups statistically. These results suggest that human umbilical cord blood CD34+ cells can be extensively expandedex vivo by using gene transfected stromal cells along with cytokines. |
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Keywords: | ex vivo expansion CD34+ cells stromal cell line Flt3 ligand thrombopoietin |
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