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Titolo:
In vitro selection of lentivirus vector-transduced human CD34(+) cells
Autore:
Gatlin, J; Douglas, J; Evans, JT; Collins, RH; Wendel, GD; Garcia, V;
Indirizzi:
Univ Texas, SW Med Ctr, Dept Internal Med, Div Infect Dis Y9 206, Dallas, TX 75235 USA Univ Texas Dallas TX USA 75235 iv Infect Dis Y9 206, Dallas, TX 75235 USA SyStem Inc, Palo Alto, CA 94304 USA SyStem Inc Palo Alto CA USA 94304SyStem Inc, Palo Alto, CA 94304 USA Univ Texas, SW Med Ctr, Dept Obstet & Gynecol, Dallas, TX 75235 USA Univ Texas Dallas TX USA 75235 ept Obstet & Gynecol, Dallas, TX 75235 USA
Titolo Testata:
HUMAN GENE THERAPY
fascicolo: 13, volume: 11, anno: 2000,
pagine: 1949 - 1957
SICI:
1043-0342(20000901)11:13<1949:IVSOLV>2.0.ZU;2-Z
Fonte:
ISI
Lingua:
ENG
Soggetto:
HUMAN HEMATOPOIETIC-CELLS; FLUORESCENT PROTEIN GENE; IMMUNE-DEFICIENT MICE; RESISTANT BONE-MARROW; UMBILICAL-CORD BLOOD; STEM-CELLS; IN-VIVO; DIHYDROFOLATE-REDUCTASE; EFFICIENT TRANSDUCTION; NUCLEOSIDE TRANSPORT;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
39
Recensione:
Indirizzi per estratti:
Indirizzo: Garcia, V Univ Texas, SW Med Ctr, Dept Internal Med, Div Infect Dis Y9 206, 5323 Harry Hines Blvd, Dallas, TX 75235 USA Univ Texas 5323 Harry Hines Blvd Dallas TX USA 75235 X 75235 USA
Citazione:
J. Gatlin et al., "In vitro selection of lentivirus vector-transduced human CD34(+) cells", HUM GENE TH, 11(13), 2000, pp. 1949-1957

Abstract

Human CD34(+) cells with in vivo repopulating potential hold much promise as a target for corrective gene transfer for numerous hematopoietic disorders. However, the efficient introduction of exogenous genes into this small,quiescent population of cells continues to present a significant challenge. To circumvent the need for high initial transduction efficiency of human hematopoietic cells, we investigated a dominant selection strategy using a variant of the DHFR gene (DHFRL22Y) For this purpose, we constructed a lentivirus-based bicistronic vector expressing EGFP and DHFRL22Y Here we demonstrate efficient in vitro selection and enrichment of lentivirus vector-transduced human CD34(+) hematopoietic cells from fetal liver, umbilical cord blood, bone marrow, and peripheral blood after cytokine mobilization, Growthof transduced human CD34(+) cells in semisolid culture under selective pressure resulted in enrichment of transduced progenitor cells to 99.5% (n = 14), Selection for DHFRL22Y+ cells after expansion of transduced progenitorsin liquid culture resulted in a 7- to 13-fold increase in the percentage of marked cells. Thus we have shown that transduced human hematopoietic cells may be effectively enriched in vitro by dominant selection, suggesting that development of such strategies holds promise for future in vivo application.

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Documento generato il 23/09/20 alle ore 11:15:06