HIGHER EFFICIENCY OF RETROVIRUS TRANSDUCTION IN THE LATE-STAGE OF PRIMARY CULTURE OF HEPATOCYTES FROM NONTREATED THAN FROM PARTIALLY HEPATECTOMIZED RAT

The aim of this investigation was to optimize the conditions for gene transduction by a retroviral vector into primary cultured hepatocytes. Because a retrovirus infection is dependent upon the proliferative ac tivity of the target cells, we examined the thymidine incorporation of the primary culture, using cells from nontreated and partially hepate ctomized donor rats. Partial hepatectomy 1 day before cell isolation g reatly enhanced the thymidine incorporation, and similarly, the transd uction of LacZ, the gene for E. coli P-galactosidase, although the per centage of transduced cells remained low, With cells from the normal r at, the thymidine incorporation increased gradually after the beginnin g of the culture and reached 16 times the initial activity on day 2, w hile hepatocytes from the partially hepatectomized donor showed no inc rease at that stage of the culture, Correspondingly, the number of gen e-transduced hepatocytes was increased when the vector was added to th e cells 1 or 2 days after isolation. Therefore, hepatocytes cultured f or 1 or 2 days after isolation are suitable for retroviral transductio n, (C) 1998 Elsevier Science Inc.