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

Proc Natl Acad Sci U S A. 1990 May;87(10):3738-42. doi: 10.1073/pnas.87.10.3738.

Abstract

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

  • Animals
  • Base Sequence
  • Cell Line
  • Cells, Cultured
  • Culture Techniques / methods
  • DNA, Viral / genetics
  • DNA, Viral / isolation & purification
  • HeLa Cells / cytology
  • Hematopoietic Stem Cells / cytology*
  • Macaca mulatta
  • Molecular Sequence Data
  • Oligonucleotide Probes
  • Polymerase Chain Reaction
  • RNA, Viral / genetics
  • RNA, Viral / isolation & purification
  • Retroviridae / genetics*
  • Retroviridae / growth & development
  • Transfection*

Substances

  • DNA, Viral
  • Oligonucleotide Probes
  • RNA, Viral