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Titolo:
Stimulation of mouse bone marrow cells with Kit ligand, FLT3 ligand, and thrombopoietin leads to efficient retrovirus-mediated gene transfer to stem cells, whereas interleukin 3 and interleukin 11 reduce transduction of short- and long-term repopulating cells
Autore:
Wognum, AW; Visser, TP; Peters, K; Bierhuizen, MFA; Wagemaker, G;
Indirizzi:
Erasmus Univ, Inst Hematol, NL-3000 DR Rotterdam, Netherlands Erasmus Univ Rotterdam Netherlands NL-3000 DR DR Rotterdam, Netherlands
Titolo Testata:
HUMAN GENE THERAPY
fascicolo: 15, volume: 11, anno: 2000,
pagine: 2129 - 2141
SICI:
1043-0342(200010)11:15<2129:SOMBMC>2.0.ZU;2-X
Fonte:
ISI
Lingua:
ENG
Soggetto:
GREEN FLUORESCENT PROTEIN; ALPHA-THALASSEMIC MICE; EX-VIVO EXPANSION; CORD BLOOD-CELLS; HEMATOPOIETIC-CELLS; PERIPHERAL-BLOOD; AUTOLOGOUS TRANSPLANTATION; RECONSTITUTING ABILITY; MONOCLONAL-ANTIBODIES; IMMUNODEFICIENT MICE;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
44
Recensione:
Indirizzi per estratti:
Indirizzo: Wognum, AW StemCell Technol Inc, 27 E 7th Ave, Vancouver, BC V5T 1M4, Canada StemCell Technol Inc 27 E 7th Ave Vancouver BC Canada V5T 1M4 a
Citazione:
A.W. Wognum et al., "Stimulation of mouse bone marrow cells with Kit ligand, FLT3 ligand, and thrombopoietin leads to efficient retrovirus-mediated gene transfer to stem cells, whereas interleukin 3 and interleukin 11 reduce transduction of short- and long-term repopulating cells", HUM GENE TH, 11(15), 2000, pp. 2129-2141

Abstract

The effects of cytokine stimulation during retroviral transduction on in vivo reconstitution of mouse hematopoietic stem cells was tested in a murinecompetitive repopulation assay with alpha-thalassemia as a marker to distinguish donor and recipient red blood cells (RBCs) and the enhanced green fluorescent protein (EGFP) as a marker for gene transfer. After transplantation, EGFP was detected in up to 90% of circulating RBCs, platelets, and leukocytes, and in primitive progenitors in bone marrow (BM), spleen, and thymus of individual transplanted mice for observation periods of more than 6 months. Large quantitative differences in reconstitution were observed after transplantation with graded numbers (1000-30,000) of EGFP(+) cells preconditioned with various combinations of Kit ligand (KL), FLT-3 ligand (FL), thrombopoietin (TPO), interleukin 3 (IL-3), and IL-11. Relative to nonmanipulated BM cells, repopulation of EGFP(+) cells was maintained by KL/FL/TPO stimulation, but, similar to 30-fold reduced after stimulation with KL/FL/TPO/IL-11, KL/FL/TPO/IL-3, or KL/FL/IL-3/IL-11. These differences were not caused by changes in the ability of immature hematopoietic cells to home to theBM, which was only moderately reduced. In conclusion, these quantitative transplantation studies of mice demonstrate the importance of optimal ex vivo cytokine stimulation for gene transfer to stem cells with retention of their in vivo hematopoietic potential, and also emphasize that overall in vitro transduction frequency does not predict gene transfer to repopulating stem cells.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 01/04/20 alle ore 23:53:59