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Infection of Hematopoietic and Stromal Ceils in Human Continuous Bone Marrow Cultures by a Retroviral Vector Containing the Neomycin Resistance Gene

Infection of Hematopoietic and Stromal Ceils in Human Continuous Bone Marrow Cultures by a... Stability and expression of the bacterial neomycin resistance gene (neo<sup>r</sup>) transferred to human continuous marrow cultures by a retroviral vector [pZIP-NeoSV(X)] was evaluated over 4 weeks. Following infection of long-term human marrow cultures with pZIP-NeoSV(X), 10-15% of the stromal cells demonstrated high replating efficiency in a dose of the neomycin analogue G418 that was toxic to stromal cells from uninfected cultures. In contrast, G418 resistance was detected in ≤1% of GM-CFUc and CFU-GEMM derived from the same virus-infected compared to control cultures. Infection of human CFU-GEMM enriched 100× by monoclonal antibody selection with pZIP-NeoSV(X) did not increase the percentage of neo<sup>r</sup> progenitors. Marrow cells from cultures infected with pZIP-NeoSV(X) and a replication competent amphotropic virus transferred the vector and G418 resistance to HeLa cells at a frequency of 1/10<sup>5</sup> for nonadherent and 1/10<sup>4</sup> for adherent cells. Two established human hematopoietic (HL60 and K562) and one stromal cell line (KM 101) stably expressed the neo<sup>r</sup> gene. Thus, a higher efficiency of infection and expression of a gene transferred by pZIP-NeoSV(X) to permanent human hematopoietic tumor cell lines and fresh marrow stromal cells contrasts with a lower level of expression in fresh CSF-dependent human hematopoietic stem cells. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Acta Haematologica Karger

Infection of Hematopoietic and Stromal Ceils in Human Continuous Bone Marrow Cultures by a Retroviral Vector Containing the Neomycin Resistance Gene

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Publisher
Karger
Copyright
© 1989 S. Karger AG, Basel
ISSN
0001-5792
eISSN
1421-9662
DOI
10.1159/000205362
Publisher site
See Article on Publisher Site

Abstract

Stability and expression of the bacterial neomycin resistance gene (neo<sup>r</sup>) transferred to human continuous marrow cultures by a retroviral vector [pZIP-NeoSV(X)] was evaluated over 4 weeks. Following infection of long-term human marrow cultures with pZIP-NeoSV(X), 10-15% of the stromal cells demonstrated high replating efficiency in a dose of the neomycin analogue G418 that was toxic to stromal cells from uninfected cultures. In contrast, G418 resistance was detected in ≤1% of GM-CFUc and CFU-GEMM derived from the same virus-infected compared to control cultures. Infection of human CFU-GEMM enriched 100× by monoclonal antibody selection with pZIP-NeoSV(X) did not increase the percentage of neo<sup>r</sup> progenitors. Marrow cells from cultures infected with pZIP-NeoSV(X) and a replication competent amphotropic virus transferred the vector and G418 resistance to HeLa cells at a frequency of 1/10<sup>5</sup> for nonadherent and 1/10<sup>4</sup> for adherent cells. Two established human hematopoietic (HL60 and K562) and one stromal cell line (KM 101) stably expressed the neo<sup>r</sup> gene. Thus, a higher efficiency of infection and expression of a gene transferred by pZIP-NeoSV(X) to permanent human hematopoietic tumor cell lines and fresh marrow stromal cells contrasts with a lower level of expression in fresh CSF-dependent human hematopoietic stem cells.

Journal

Acta Haematologica – Karger

Published: Jan 1, 1989

Keywords: Gene expression; Gene therapy; Human marrow stroma

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