Efficient in vivo xenogeneic retroviral vector-mediated gene transduction into human hepatocytes.
We developed a method for efficient retroviral vector-mediated gene transfer into human hepatocytes, using a human hepatocyte-bearing mouse model. Normal human hepatocytes were transplanted into the livers of immunodeficient and liver-damaged mice. Donor hepatocytes multiplied and replaced the host hepatocytes, which yielded human hepatocyte-bearing mice (human hepatocyte-chimeric mice). As control cells, rat hepatocytes were similarly transplanted. The replacement level reached 86% at 8 weeks and 100% at 5 weeks posttransplantation of human and rat hepatocytes, respectively. Human and rat hepatocytes in the host liver showed a high bromodeoxyuridine-labeling index during the first 2 weeks posttransplantation. Human- and rat-chimeric mice were injected 7 and 10 days posttransplantation, respectively, with retroviral vectors carrying the beta-galactosidase gene and were thereafter injected daily for 20 and 10 days, respectively. The level of beta-galactosidase-positive hepatocytes in the human- and rat-chimeric mice reached 7.1 +/- 1.8% at 8 weeks and 5.3 +/- 0.9% at 5 weeks after transplantation, respectively. The human hepatocyte-chimeric mouse will be useful for testing the ability of vectors to transduce human cells.
Emoto
Kentaro
K
Yoshizato Project, Cooperative Link of Unique Science and Technology for Economy Revitalization (CLUSTER), Hiroshima Prefectural Institute of Industrial Science and Technology, Higashihiroshima, Hiroshima 739-0046, Japan.
Tateno
Chise
C
Hino
Hiroshi
H
Amano
Hironobu
H
Imaoka
Yasuhiro
Y
Asahina
Kinji
K
Asahara
Toshimasa
T
Yoshizato
Katsutoshi
K
D000818:Animals; D015316:Genetic Therapy / Q000379:methods; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism / Q000637:transplantation; D006801:Humans; D051379:Mice; D008817:Mice, Mutant Strains; D051381:Rats; D012190:Retroviridae / Q000235:genetics; D014161:Transduction, Genetic / Q000379:methods; D018183:Transplantation Chimera / Q000235:genetics / Q000378:metabolism; D014183:Transplantation, Heterologous