A stable gene transfer system for hematopoietic progenitor cells from human bone marrow using pseudotyped retroviral vectors

Sang Won Park, Sang Yun Choi

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Recombinant DNA technology has permitted tremendous progression in delivering genes into cells; however, further advances in gene replacement techniques are needed prior to application to hematological diseases. One of the greatest obstacles to gene therapy in human hematopoietic stem cells is the lack of a defined protocol in humans and low transduction efficiency. Currently, murine leukemia virus (MuLV) is the most popular choice as a gene transfer vehicle but it cannot infect non-dividing cells. In our study, vesicular stomatitis G protein pseudotyped MuLV and HIV-1 were produced by a split gene transfection method. Mono-nuclear cells were separated from healthy human bone marrow and pre-stimulated with cytokines to form myeloid cell lineages. The cells were infected at different MOIs with highly concentrated virus and infection rates were analyzed by flow cytometry and progenitor cell assays. eGFP expression was much higher when using HIV-1 system than when using MuLV. Progenitor cell assays agreed with the results obtained by FACS, but the difference was less great. We conclude that the lentiviral system is more suitable for gene transfer to hematopoietic progenitor cells probably because it stably infects both dividing and non-dividing cells. In addition, fibronectin was shown to improve the rate of infection with HIV-1.

Original languageEnglish
Pages (from-to)297-303
Number of pages7
JournalMolecules and cells
Volume17
Issue number2
Publication statusPublished - 2004 Apr 30

Keywords

  • Gene therapy
  • Hematopoietic progenitor cells
  • Lentivirus
  • Murine leukemia virus

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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