Simplified production and concentration of lentiviral vectors to achieve high transduction in primary human T cells.
BMC Biotechnol
; 13: 98, 2013 Nov 12.
Article
en En
| MEDLINE
| ID: mdl-24215295
ABSTRACT
BACKGROUND:
Lentiviral vectors have emerged as efficient vehicles for transgene delivery in both dividing and non-dividing cells. A number of different modifications in vector design have increased biosafety and transgene expression. However, despite these advances, the transduction of primary human T cells is still challenging and methods to achieve efficient gene transfer are often expensive and time-consuming.RESULTS:
Here we present a simple optimised protocol for the generation and transduction of lentivirus in primary human CD45RA+ T cells. We show that generation of high-titre lentivirus with improved primary T cell transduction is dependent upon optimised ultracentrifuge speed during viral concentration. Moreover, we demonstrate that transduction efficiency can be increased with simple modifications to the culturing conditions. Overall, a transduction efficiency of up to 89% in primary human CD45RA+ cells is achievable when these modifications are used in conjunction.CONCLUSION:
The optimised protocol described here is easy to implement and should facilitate the production of high-titre lentivirus with superior transduction efficiency in primary human T cells without the need for further purification methods.
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Linfocitos T
/
Lentivirus
/
Vectores Genéticos
Límite:
Humans
Idioma:
En
Revista:
BMC Biotechnol
Asunto de la revista:
BIOTECNOLOGIA
Año:
2013
Tipo del documento:
Article
País de afiliación:
Reino Unido