GaLV Pseudotyped Vectors and Cationic Lipids Transduce Human CD34+Cells

Abstract

High transduction frequency of hematopoietic stem/progenitor cells is essential to derive clinical benefits for treating certain inherited and acquired diseases. We demonstrate here stable gene transfer into human bone marrow-derived CD34⁺ progenitors using cationic lipids to facilitate GaLV and amphotroic pseudotyped retroviral-mediated transductions. Furthermore, the transgene was detected only in the progeny of flow cytometer sorted CD34⁺ population transduced by the LAPSN (PG13) viral vector in the presence of cationic lipids but not when transduction was facilitated with conventional polycations Polybrene or protamine sulfate. Thus, a combination of GaLV pseudotyped vectors and cationic lipids results in increased transduction frequencies of the CD34⁺ cells without a requirement of extended in vitro culture, or co-cultivation with producer cell lines. These improvements may result in the production of therapeutically significant quantities of genetically modified hematopoietic cells. A protocol using retroviral vectors was optimized for gene transfer into human bone marrow progenitor cells. The transduction frequencies observed for human CD34⁺ cells using GaLV pseudotyped retroviral vector and cationic lipids were significantly higher than those obtained with routinely used combinations of amphotropic pseudotype of the same vector and Polybrene or protamine sulfate. Additionally, transduction frequency of human CD34⁺ cells was higher when infected with a combination of GaLV pseudotyped vector and cationic lipids as compared to the combination of amphotropic pseudotyped vector and cationic lipids. These increased transduction frequencies may improve the potential clinical utility of certain gene therapies using transduced hematopoietic stem/progenitor cells.