The Stimulatory Effect of MIP-1α on the Gene Transduction Efficacy of Cord Blood CD34+ Cells by a Pseudotype Retroviral Vector
-
Kamran Alimoghaddam
,
L Moezzi
,
Hamid Ghaffari
,
Alireza Arjmand
,
P Ghodsi
,
Bahram Chahardouli
,
S Shahrokhi
,
Ardeshir Ghavamzadeh
Abstract
Introduction: Hematopoietic stem cells are always in a quiescence state. Since they need retroviral transduction to infect dividing cells, they are resistant to retrovirus transduction. They need to be pre-stimulated by a cytokine cocktail. For proliferation without maturation, we suggest MIP-1α as a novel factor.
Material and methods: Retroviral vector produced by PG13/LN C8 cells titter on Hela cells. Then, the CD34+ cells of cord blood can be pre-stimulated in a serum- free media supplemented with SCF, Flt3,TPO,IL6 in the presence and absence of 50 ng/ml MIP-1α. Transduction efficiency was assessed by a semi-quantitative PCR for the neomycin gene.
Results: A PCR analysis of the neomycin gene in CD34+ cells revealed an improved transduction of cord blood cells in the presence of MIP-1α 65%, in comparison to its absence: 40.7%.
Conclusion: the addition of MIP-1α to the cytokine cocktail improves the transduction efficiency of cord blood hematopoietic progenitor cells. Further studies are required to clarify its effect on the functional properties of CD34+ cells.
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| Issue | Vol 3, No 2 (2009) | |
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