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Journal of Cell Science, Vol 99, Issue 1 121-130, Copyright © 1991 by Company of Biologists

JOURNAL ARTICLES

Retroviral vector infection and transplantation in rats of primary fetal rat hepatocytes

KS Koch, GG Brownlee, SJ Goss, A Martinez-Conde and HL Leffert
Department of Pharmacology, University of California, San Diego, La Jolla.

Retroviral vectors were used to transduce recombinant DNA encoding firefly luciferase, Escherichia coli beta-galactosidase or human factor IX into fetal rat hepatocytes in primary culture. Hepatocytes were transduced optimally during a restricted time interval, 2-4 days post-plating. Although efficient and stable expression of reporter gene products was observed in vitro, it was affected differentially by culture conditions (plating density, media constituents) and chemical modulators of hepatocyte growth and differentiation (gelatin, hydrocortisone, isobutylmethylxanthine). Cultured cells, mock-infected or infected with a luciferase-expressing vector, were harvested non-enzymatically and injected subcutaneously into the dorsal neck fascia of neonatal syngeneic rats. Tissue isolated from injection sites one week later contained hepatocyte foci. In animals transplanted with infected cells, the preliminary results suggest that luciferase activity was present at these sites in proportion to the numbers of injected cells. These findings and previous observations made with hepatocytes from neonatal and adult primary cultures, indicate that from day 19 in utero through maturity the transient temporal 'period of susceptibility' to infection in vitro is independent of the developmental state of starting tissue. Transplantability of cultured fetal hepatocytes infected with retroviral vectors and stably expressing reporter gene products suggests that such cells might provide promising models for liver gene therapy.


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E. D. Zanjani and W. F. Anderson
Prospects for in Utero Human Gene Therapy
Science, September 24, 1999; 285(5436): 2084 - 2088.
[Abstract] [Full Text]


© The Company of Biologists Ltd 1991