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Improved Coinfection with Amphotropic Pseudotyped Retroviral Vectors
Joint Authors
Zhang, Yong
Hornsby, Peter J.
Melton, David W.
Wu, Yuehong
Source
Journal of Biomedicine and Biotechnology
Issue
Vol. 2009, Issue 2009 (31 Dec. 2009), pp.1-7, 7 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2009-05-25
Country of Publication
Egypt
No. of Pages
7
Main Subjects
Abstract EN
Amphotropic pseudotyped retroviral vectors have typically been used to infect target cells without prior concentration.
Although this can yield high rates of infection, higher rates may be needed where highly efficient coinfection of two or more vectors is needed.
In this investigation we used amphotropic retroviral vectors produced by the Plat-A cell line and studied coinfection rates using green and red fluorescent proteins (EGFP and dsRed2).
Target cells were primary human fibroblasts (PHF) and 3T3 cells.
Unconcentrated vector preparations produced a coinfection rate of ∼4% (defined as cells that are both red and green as a percentage of all cells infected).
Optimized spinoculation, comprising centrifugation at 1200 g for 2 hours at 15∘C, increased the coinfection rate to ∼10%.
Concentration by centrifugation at 10,000 g or by flocculation using Polybrene increased the coinfection rate to ∼25%.
Combining the two processes, concentration by Polybrene flocculation and optimized spinoculation, increased the coinfection rate to 35% (3T3) or >50% (PHF).
Improved coinfection should be valuable in protocols that require high transduction by combinations of two or more retroviral vectors.
American Psychological Association (APA)
Wu, Yuehong& Melton, David W.& Zhang, Yong& Hornsby, Peter J.. 2009. Improved Coinfection with Amphotropic Pseudotyped Retroviral Vectors. Journal of Biomedicine and Biotechnology،Vol. 2009, no. 2009, pp.1-7.
https://search.emarefa.net/detail/BIM-506518
Modern Language Association (MLA)
Wu, Yuehong…[et al.]. Improved Coinfection with Amphotropic Pseudotyped Retroviral Vectors. Journal of Biomedicine and Biotechnology No. 2009 (2009), pp.1-7.
https://search.emarefa.net/detail/BIM-506518
American Medical Association (AMA)
Wu, Yuehong& Melton, David W.& Zhang, Yong& Hornsby, Peter J.. Improved Coinfection with Amphotropic Pseudotyped Retroviral Vectors. Journal of Biomedicine and Biotechnology. 2009. Vol. 2009, no. 2009, pp.1-7.
https://search.emarefa.net/detail/BIM-506518
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-506518