title: 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.
authors.affiliation: Targeted Genetics Corporation, Seattle, WA 98101, USA.
authors.forename: J
authors.initials: J
authors.lastname: Dybing
authors.forename: C M
authors.initials: CM
authors.lastname: Lynch
authors.forename: P
authors.initials: P
authors.lastname: Hara
authors.forename: L
authors.initials: L
authors.lastname: Jurus
authors.forename: H P
authors.initials: HP
authors.lastname: Kiem
authors.forename: P
authors.initials: P
authors.lastname: Anklesaria
mesh_terms: D000469:Alkaline Phosphatase / Q000235:genetics; D018952:Antigens, CD34 / Q000032:analysis*; D001854:Bone Marrow Cells; D002412:Cations; D002460:Cell Line; D002478:Cells, Cultured; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D017930:Genes, Reporter / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D006412:Hematopoietic Stem Cells / Q000737:chemistry; D006583:Hexadimethrine Bromide; D006801:Humans; D017989:Leukemia Virus, Gibbon Ape / Q000235:genetics*; D008055:Lipids; D008979:Moloney murine leukemia virus / Q000235:genetics; D013096:Spermine / Q000031:analogs & derivatives
chemical_list: C098223:2,3-dioleyloxy-N-(2-(sperminecarboxamido)ethyl)-N,N-dimethyl-1-propanaminium trifluoroacetate; D018952:Antigens, CD34; D002412:Cations; D008055:Lipids; D013096:Spermine; D006583:Hexadimethrine Bromide; D000469:Alkaline Phosphatase