Novel retroviral packaging cell lines: complementary tropisms and improved vector production for efficient gene transfer (original) (raw)
- Paper
- Published: 01 June 1997
Gene Therapy volume 4, pages 600–610 (1997)Cite this article
- 799 Accesses
- 35 Citations
- 3 Altmetric
- Metrics details
Abstract
We report increased transduction of human hematopoietic progenitor cells through a combination of novel retroviral vector packaging cell lines, and improved vector supernatant production. The new ProPak packaging cell lines produce either murine leukemia virus (MLV) xenotropic (ProPak-X cells) or amphotropic particles (ProPak-A cells), and ProPak-based producer cells were demonstrated to be free of replication-competent retrovirus (RCR) by stringent testing. Vector supernatants from ProPak or existing packaging cell lines producing different pseudotyped particles (amphotropic MLV, xenotropic MLV or gibbon ape leukemia virus) were compared for the ability to transduce clinically relevant human hematopoietic cells. All vector types transduced primary human CD34-positive or CD4-positive cells, regardless of tropism. However, consistently higher transduction of target cells was achieved with ProPak-derived amphotropic vector than with PA317-packaged amphotropic vector. The highest transduction of human hematopoietic progenitor cells was achieved with vector supernatant generated from a coculture of the ProPak-X and ProPak-A cell lines. This ping-pong amplification yielded supernatant containing vector targeted to two distinct receptors present on human cells, and did not result in detectable RCR formation. In addition, we describe conditions for improved vector supernatant production in a packed-bed bioreactor.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on SpringerLink
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Additional access options:
Similar content being viewed by others
Author information
Authors and Affiliations
- Progenesys Program, SyStemix Inc., 3155 Porter Drive, Palo Alto, 94304, CA, USA
SP Forestell, JS Dando, J Chen, P de Vries, E Böhnlein & RJ Rigg - Gene Therapy Programme, DIBIT HS Raffaele, Milan, Italy
JS Dando - Cell Therapeutics, Seattle, Washington, USA
P de Vries
Authors
- SP Forestell
You can also search for this author inPubMed Google Scholar - JS Dando
You can also search for this author inPubMed Google Scholar - J Chen
You can also search for this author inPubMed Google Scholar - P de Vries
You can also search for this author inPubMed Google Scholar - E Böhnlein
You can also search for this author inPubMed Google Scholar - RJ Rigg
You can also search for this author inPubMed Google Scholar
Rights and permissions
About this article
Cite this article
Forestell, S., Dando, J., Chen, J. et al. Novel retroviral packaging cell lines: complementary tropisms and improved vector production for efficient gene transfer.Gene Ther 4, 600–610 (1997). https://doi.org/10.1038/sj.gt.3300420
- Received: 22 October 1996
- Accepted: 27 January 1997
- Issue Date: 01 June 1997
- DOI: https://doi.org/10.1038/sj.gt.3300420