Abstract
Cultured human BMC have been infected with a transmissible amphotrophic murine retroviral vector containing the human HPRT cDNA. Infection of the cultured cells and production of progeny vector has been determined by a colony formation assay in which culture medium from the infected cells is used to infect HPRT-deficient mouse fibroblasts. Successful marrow cell infection demonstrated by progeny virus production is indicated by formation of colonies of mouse cells resistant to hypoxanthine-aminopterin-thymidine (HAT) selective medium. Infected bulk marrow cells have been found to produce detectable progeny HPRT virus, and the conditions of infection including such variables as multiplicity of infection, time of infection, serum conditions, polybrene concentration and others are being optimized. Successful infection of a stem cell population has been demonstrated by efficient virus production from isolated granulocyte/monocyte colony forming units (CFU-GM) growing in semisolid (agar) medium.
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Gruber, H., Friedmann, T., Finley, K. et al. INSERTION OF HYPOXAKTHINE PHOSPHORIBOSYLTRANSFERASE (HPRT) cDNA INTO HUMAN BONE MARROW CELLS (BMC) A RETROVIRUS VECTOR: 75. Pediatr Res 19, 756 (1985). https://doi.org/10.1203/00006450-198507000-00095
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DOI: https://doi.org/10.1203/00006450-198507000-00095