scholarly journals Generation of a stable packaging cell line producing high-titer PPT-deleted integration-deficient lentiviral vectors

2015 ◽  
Vol 2 ◽  
pp. 15025 ◽  
Author(s):  
Peirong Hu ◽  
Yedda Li ◽  
Mark S Sands ◽  
Thomas McCown ◽  
Tal Kafri
Keyword(s):  
Cell Line ◽  
High Titer ◽  
Lentiviral Vectors ◽  
Packaging Cell Line ◽  
Packaging Cell

Highly efficient gene transfer into baboon marrow repopulating cells using GALV-pseudotype oncoretroviral vectors produced by human packaging cells

Blood ◽  
2002 ◽  
Vol 100 (12) ◽  
pp. 3960-3967 ◽  
Author(s):  
Peter A. Horn ◽  
Max S. Topp ◽  
Julia C. Morris ◽  
Stanley R. Riddell ◽  
Hans-Peter Kiem
Keyword(s):  
Stem Cells ◽  
Gene Transfer ◽  
Cell Line ◽  
Flow Cytometric Analysis ◽  
Lentiviral Vectors ◽  
Transduction Efficiency ◽  
Packaging Cell Line ◽  
Hematopoietic Stem ◽  
Packaging Cell ◽  
Cd34 Cells

Vector-containing medium harvested from murine packaging cell lines has been shown to contain factors that can negatively influence the transduction and maintenance of hematopoietic stem cells. Thus, we generated a human packaging cell line with a gibbon ape leukemia virus pseudotype (Phoenix-GALV), and we evaluated vectors produced by Phoenix-GALV for their ability to transduce hematopoietic progenitor/stem cells. In 3 baboons, we used a competitive repopulation assay to directly compare GALV-pseudotype retrovirus vectors produced by either Phoenix-GALV or by the NIH 3T3–derived packaging cell line, PG13. In 3 additional baboons we compared Phoenix-GALV–derived vectors to more recently developed lentiviral vectors. Gene transfer efficiency into hematopoietic repopulating cells was assessed by evaluating the number of genetically modified peripheral blood and marrow cells using flow cytometry and real-time polymerase chain reaction. Transduction efficiency of hematopoietic repopulating cells was significantly higher using the Phoenix-GALV–derived vector as compared with the PG13-derived vectors or lentiviral vectors, with stable transduction levels up to 25%. We followed 2 animals for more than one year. Flow cytometric analysis of hematopoietic subpopulations in these animals revealed transgene expression in CD13+ granulocytes, CD20+ B lymphocytes, CD3+ T lymphocytes, CD61+ platelets, as well as red blood cells, indicating multilineage engraftment of cells transduced by Phoenix-GALV–pseudotype vectors. In addition, transduction of human CD34+ cells was significantly more efficient than transduction of baboon CD34+ cells, suggesting that Phoenix-GALV–derived oncoretroviral vectors may be even more efficient in human stem cell gene therapy applications.

scholarly journals A trans-lentiviral packaging cell line for high-titer conditional self-inactivating HIV-1 vectors

2006 ◽  
Vol 14 (2) ◽  
pp. 276-284 ◽  
Author(s):  
Adam S. Cockrell ◽  
Hong Ma ◽  
Kailing Fu ◽  
Thomas J. McCown ◽  
Tal Kafri
Keyword(s):  
Cell Line ◽  
High Titer ◽  
Packaging Cell Line ◽  
Packaging Cell ◽  
Hiv 1

scholarly journals RD2-MolPack-Chim3,a Packaging Cell Line for Stable Production of Lentiviral Vectors for Anti-HIV Gene Therapy

2013 ◽  
Vol 24 (4) ◽  
pp. 228-240 ◽  
Author(s):  
Anna Stornaiuolo ◽  
Bianca Maria Piovani ◽  
Sergio Bossi ◽  
Eleonora Zucchelli ◽  
Stefano Corna ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Cell Line ◽  
Lentiviral Vectors ◽  
Packaging Cell Line ◽  
Packaging Cell ◽  
Stable Production ◽  
Anti Hiv

scholarly journals A Packaging Cell Line for Lentivirus Vectors

1999 ◽  
Vol 73 (1) ◽  
pp. 576-584 ◽  
Author(s):  
Tal Kafri ◽  
Henriette van Praag ◽  
Ling Ouyang ◽  
Fred H. Gage ◽  
Inder M. Verma
Keyword(s):  
Cell Line ◽  
High Titer ◽  
Packaging Cell Line ◽  
Virus Particles ◽  
Packaging Cell ◽  
Retinal Tissue ◽  
Lentivirus Vectors ◽  
Inducible Cell Line

ABSTRACT Lentivirus vectors can transduce dividing and nondividing cells. Using three-plasmid transient transfections, high-titer (>109 IU/ml) recombinant lentivirus vectors pseudotyped with vesicular stomatitis virus G (VSV-G) protein can be generated (T. Kafri et al., Nat. Genet. 17:314–317, 1997; H. Miyoshi et al., Proc. Natl. Acad. Sci. USA 94:10319–10323, 1997; L. Naldini et al., Science 272:263–267, 1996). The recombinant lentiviruses can efficiently infect brain, liver, muscle, and retinal tissue in vivo. Furthermore, the transduced tissues demonstrated long-term expression of reporter genes in immunocompetent rodents. We now report the generation of a tetracycline-inducible VSV-G pseudotyped lentivirus packaging cell line which can generate virus particles at titers greater than 106 IU/ml for at least 3 to 4 days. The vector produced by the inducible cell line can be concentrated to titers of 109 IU/ml and can efficiently transduce nondividing cells in vitro and in vivo. The availability of a lentivirus packaging cell line will significantly facilitate the production of high-titer lentivirus vectors for gene therapy and study of human immunodeficiency virus biology.

A New-Generation Stable Inducible Packaging Cell Line for Lentiviral Vectors

2001 ◽  
Vol 12 (8) ◽  
pp. 981-997 ◽  
Author(s):  
Deborah Farson ◽  
Rochelle Witt ◽  
Ryan McGuinness ◽  
Tom Dull ◽  
Michael Kelly ◽  
...  
Keyword(s):  
Cell Line ◽  
Lentiviral Vectors ◽  
Packaging Cell Line ◽  
Packaging Cell ◽  
New Generation
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