Retrovirus-mediated transfer of a hygromycin phosphotransferase-thymidine kinase fusion gene into human CD34+ bone marrow cells

Yoshiki Akatsuka, Nobuhiko Emi, Hidefumi Kato, Akihiro Abe, Mitsune Tanimoto, Stephen D. Lupton, Hidehiko Saito

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Retrovirus-mediated gene transfer into human hematopoietic stem cells has been proposed as a means of therapy for various inherited diseases and as a method of gene marking. The transduction efficiency of an amphotropic retroviral vector (PA317/HyTK) containing a hygromycin phosphotransferase-thymidine kinase fusion gene was examined with human CD34+ bone marrow cells in the presence of interleukin-3 (IL-3), interleukin-6 (IL-6), and stem cell factor. Transduction efficiencies determined from the ability of transduced granulocyte-macrophage colony forming units (CFU-GM) to grow in hygromycin B and from polymerase chain reaction analysis of individual transduced CFU-GM growing in the presence of hygromycin B were 0.3-3.0% (mean ± S.D., 1.1 ± 0.9%) and 0.1-1.2% (mean ± S.D., 0.5 ± 0.4%), respectively. Ganciclovir at a dose of ∼ 1 μM reduced the number of CFU-GM derived from vector-infected CD34+ cells by 50%. These findings demonstrate that human hematopoietic stem cells infected with this retroviral vector are susceptible to ganciclovir, offering the potential to control transduced gene expression in vivo.

Original languageEnglish
Pages (from-to)251-261
Number of pages11
JournalInternational Journal of Hematology
Issue number4
Publication statusPublished - 01-12-1994


All Science Journal Classification (ASJC) codes

  • Hematology

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