Development of a high-titer retrovirus producer cell line capable of gene transfer into rhesus monkey hematopoietic stem cells.

Bodine DM, McDonagh KT, Brandt SJ, Ney PA, Agricola B, Byrne E, Nienhuis AW
Proc Natl Acad Sci U S A. 1990 87 (10): 3738-42

PMID: 2339117 · PMCID: PMC53978 · DOI:10.1073/pnas.87.10.3738

Retroviral-mediated gene transfer into primitive hematopoietic cells has been difficult to achieve in large-animal models. We have developed an amphotropic producer clone that generates greater than 10(10) recombinant retroviral particles (colony-forming units) per ml of culture medium. Autologous rhesus monkey bone-marrow cells were cocultured with either high (2 x 10(10) colony-forming units/ml) or low (5 x 10(6) colony-forming units/ml) titer producer clones for 4-6 days and reinfused into sublethally irradiated animals. The proviral genome was detected in blood and bone-marrow cells from all three animals reconstituted with cells cocultured with the high-titer producer cells. In contrast, three animals reconstituted with bone marrow cocultured with the low-titer producer clone exhibited no evidence of gene transfer.

MeSH Terms (15)

Animals Base Sequence Cell Line Cells, Cultured Culture Techniques DNA, Viral HeLa Cells Hematopoietic Stem Cells Macaca mulatta Molecular Sequence Data Oligonucleotide Probes Polymerase Chain Reaction Retroviridae RNA, Viral Transfection

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