Optimized retroviral transduction protocol which preserves the primitive subpopulation of human hematopoietic cells

Tonks, Alex ORCID: https://orcid.org/0000-0002-6073-4976, Tonks, Amanda Jayne ORCID: https://orcid.org/0000-0002-6073-4976, Pearn, Lorna, Mohamad, Zulkhairi, Burnett, Alan Kenneth and Darley, Richard Lawrence ORCID: https://orcid.org/0000-0003-0879-0724 2005. Optimized retroviral transduction protocol which preserves the primitive subpopulation of human hematopoietic cells. Biotechnology Progress 21 (3) , pp. 953-958. 10.1021/bp0500314

Full text not available from this repository.

Abstract

Though both low-speed centrifugation and the use of fibronectin (Retronectin) fragments increase gene transduction efficiency, they still do not overcome the adverse effects of the presence of virus-containing medium (VCM). In this study, we improved transduction efficiency of primitive human hematopoietic cells by optimizing the conditions for preadsorbing culture dishes with retrovirus using a centrifugation protocol allowing subsequent infection to be carried out in the absence of VCM. We also demonstrate that preadsorbing tissue culture plates with retrovirus is dependent on the volume of VCM used for preadsorption and the length of centrifugation and the type of plasticware used but not on the temperature of centrifugation (4–33 °C). Direct exposure of CD34+ target cells to VCM depletes the primitive CD34+CD38neg subpopulation by more than 30%, whereas the optimized VCM-free infection protocol targets this population with equivalent efficiency but had no detrimental effects on CD34+CD38neg frequency. In summary, we demonstrate a high-frequency transduction protocol which preserves the therapeutically relevant primitive subpopulation of human hematopoietic cells.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Medicine
Subjects:	R Medicine > R Medicine (General)
Publisher:	John Wiley & Sons
ISSN:	8756-7938
Last Modified:	25 Oct 2022 09:26
URI:	https://orca.cardiff.ac.uk/id/eprint/58482

Citation Data

Cited 19 times in Scopus. View in Scopus. Powered By Scopus® Data

Actions (repository staff only)

Edit Item