Catalogo Articoli (Spogli Riviste)

OPAC HELP

Titolo:
Retroviral transduction of human CD34(+) cells on fibronectin fragment CH-296 is inhibited by high concentrations of vector containing medium
Autore:
Relander, T; Brun, A; Hawley, RG; Karlsson, S; Richter, J;
Indirizzi:
Univ Lund, Wallenberg Neurosci ctr, Dept Mol Med & Gene Therapy, S-22362 Lund, Sweden Univ Lund Lund Sweden S-22362 l Med & Gene Therapy, S-22362 Lund, Sweden Amer Red Cross, Jerome H Holland Lab, Hematopoiesis Dept, Rockville, MD 20855 USA Amer Red Cross Rockville MD USA 20855 iesis Dept, Rockville, MD 20855 USA
Titolo Testata:
JOURNAL OF GENE MEDICINE
fascicolo: 3, volume: 3, anno: 2001,
pagine: 207 - 218
SICI:
1099-498X(200105/06)3:3<207:RTOHCC>2.0.ZU;2-8
Fonte:
ISI
Lingua:
ENG
Soggetto:
APE LEUKEMIA-VIRUS; HEMATOPOIETIC PROGENITOR CELLS; VESICULAR STOMATITIS-VIRUS; CULTURE-INITIATING CELLS; RECEPTOR MESSENGER-RNA; MEDIATED GENE-TRANSFER; LONG-TERM CULTURES; PERIPHERAL-BLOOD; BONE-MARROW; HIGH-TITER;
Keywords:
gene transfer; retroviral; CD34(+); NOD/SCID; cord blood;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
43
Recensione:
Indirizzi per estratti:
Indirizzo: Richter, J Univ Lund, Wallenberg Neurosci ctr, Dept Mol Med & Gene Therapy, Solvegatan 17, S-22362 Lund, Sweden Univ Lund Solvegatan 17 Lund Sweden S-22362 22362 Lund, Sweden
Citazione:
T. Relander et al., "Retroviral transduction of human CD34(+) cells on fibronectin fragment CH-296 is inhibited by high concentrations of vector containing medium", J GENE MED, 3(3), 2001, pp. 207-218

Abstract

Background The objective of the present study was to optimize conditions for retroviral transduction of human cord blood (CB) CD34(+) cells and to reveal mechanisms which interfere with efficient gene transfer. Methods An MSCV based retroviral vector with the gene for enhanced green fluorescent protein (MGIN) produced by GP + envAM12 Camphotropic envelope), PG13 (gibbon ape leukemia virus envelope) or 293GPG (vesicular stomatitis virus envelope) cell lines was used to transduce cord blood CD34(+) cells onRetronectin (fibronectin fragment CH-296) in three different ways: either in vector containing medium (VCM), in fresh medium on Retronectin pre-loaded with vector or in VCM on Retronectin pre-loaded with vector. Results Paradoxically, the transduction efficiency obtained with pre-load of vector onto Retronectin alone was higher than pre-load plus VCM for PG13-MGIN (67.9 +/- 6.0% vs 24.9 +/- 8.0%) and AM12-MGIN C47.5 +/- 5.8% vs 38.7+/- 2.2%). Further experiments showed that transduction on Retronectin pre-loaded with PG13-MGIN or AM12-MGIN was inhibited by the presence of the same VCM at high concentrations, but not by the presence of a VCM with a different receptor specificity. If no pre-load of vector was performed, the highest transduction efficiencies were seen when VCMs were diluted 1:10 (MOIs of 3). The inhibitory effect of high titer PG13-MGIN VCM was confirmed in more primitive CD34(+)CD38(low) cells and in NOD/SCID repopulating cells, and was also seen in experiments with bone marrow CD34(+) cells. Conclusions Retroviral transduction of CB CD34+ cells on Retronectin is inhibited by high titer PG13 and GP +envAM12 vector containing medium. Efficient gene transfer to human hematopoietic cells can be obtained by preload alone of the vector onto Retronectin. These findings are of importance for the design of transduction protocols for repopulating hematopoietic cells. Copyright (C) 2001 John Wiley & Sons, Ltd.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 19/01/20 alle ore 12:29:25