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Titolo:
Co-transduction of cDNAs for c-kit and steel factor into single CD34(+) cord blood cells further enhances the growth of erythroid and multipotential progenitors
Autore:
Lu, L; Dai, MS; Ge, Y; Wang, LS; Braun, SE; Wait, CL; Griffith, DJ; Heinrich, MC; Broxmeyer, HE;
Indirizzi:
Indiana Univ, Sch Med, Walther Oncol Ctr, Indianapolis, IN 46202 USA Indiana Univ Indianapolis IN USA 46202 ol Ctr, Indianapolis, IN 46202 USA Indiana Univ, Sch Med, Dept Microbiol & Immunol, Indianapolis, IN 46202 USA Indiana Univ Indianapolis IN USA 46202 mmunol, Indianapolis, IN 46202 USA Indiana Univ, Sch Med, Dept Med Hematol Oncol, Indianapolis, IN 46202 USA Indiana Univ Indianapolis IN USA 46202 Oncol, Indianapolis, IN 46202 USA Walther Canc Inst, Indianapolis, IN USA Walther Canc Inst Indianapolis INUSA er Canc Inst, Indianapolis, IN USA Oregon Hlth Sci Univ, Dept Med, Div Hematol & Med Oncol, Portland, OR 97201 USA Oregon Hlth Sci Univ Portland OR USA 97201 Oncol, Portland, OR 97201 USA Portland Vet Affairs Med Ctr, Portland, OR USA Portland Vet Affairs Med Ctr Portland OR USA s Med Ctr, Portland, OR USA
Titolo Testata:
JOURNAL OF HEMATOTHERAPY & STEM CELL RESEARCH
fascicolo: 6, volume: 9, anno: 2000,
pagine: 813 - 825
SICI:
1525-8165(200012)9:6<813:COCFCA>2.0.ZU;2-F
Fonte:
ISI
Lingua:
ENG
Soggetto:
MEDIATED GENE TRANSDUCTION; RECEPTOR TYROSINE KINASE; HUMAN-ERYTHROPOIETIN RECEPTOR; COLONY FORMATION; TRANSFORMING GROWTH-FACTOR-BETA-1; SIGNAL-TRANSDUCTION; LIGAND LACKING; STROMAL CELLS; SI-LOCUS; V-KIT;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Clinical Medicine
Life Sciences
Citazioni:
36
Recensione:
Indirizzi per estratti:
Indirizzo: Lu, L Indiana Univ, Sch Med, Walther Oncol Ctr, 1044 W Walnut St,Room 302,Indianapolis, IN 46202 USA Indiana Univ 1044 W Walnut St,Room 302 Indianapolis IN USA 46202 USA
Citazione:
L. Lu et al., "Co-transduction of cDNAs for c-kit and steel factor into single CD34(+) cord blood cells further enhances the growth of erythroid and multipotential progenitors", J HEMATH ST, 9(6), 2000, pp. 813-825

Abstract

Previous studies have demonstrated that the c-kit encoded tyrosine kinase receptor and its ligand, steel factor (SLF), are critical for normal blood cell development. We have reported that transduction of the c-kif gene intosingle hematopoietic progenitor cells (HPC), CD34(+++) cells, from cord blood (CB) enhances erythroid colony formation via a SLF-dependent mechanism. We therefore decided to evaluate the impact on cell proliferation of co-transducing c-kif and SLF cDNAs into these cells. CD34(+++) cells were sortedas a population or as 1 cell/well for cells expressing the highest levels of CD34 and different levels of c-kit. Cells were then prestimulated with granulocyte macrophage (GM)-colony stimulating factor (CSF), interleukin (IL)-3, IL-6, erythropoietin (Epo) in the presence and absence of various concentrations of SLF. Cells were then transduced with SLF and/or c-kit cDNAs, and then assayed for colony formation with the same cytokine combination. At a single cell level, co-transduction with c-kif and SLF genes significantly enhanced colony formation compared with individual gene transduction, especially by erythroid and multipotential progenitors that responded to stimulation by added cytokines. Little or no growth was seen with the c-kit- and/or SLF-transduced cells without addition of cytokines. The degree of enhancement effected by co-transduction inversely correlated with the degree ofexpression of c-kit protein before transduction. Optimal enhancing effectswere noted in CD34(+++) kit(Lo/-) cells co-transduced with both c-kit and SLF cDNAs. Reverse transcriptase-polymerase chain (RT-PCR) analysis of SLF mRNA expression in CD34(+++) cells and enzyme-linked immunoadsorbent assay (ELISA) measurement of secreted SLF protein demonstrated that the transduced SLF cDNA was expressed and soluble SLF was released in medium cultured with SLP gene transduced MACS-separated CD34(+) cells in the presence, but not in the absence, of IL-3, GM-CSF, IL-6, and Epo. These results demonstratethe enhancement of the proliferation of growth factor responsive HPC that express transduced c-kit and SLF genes.

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Documento generato il 01/04/20 alle ore 23:57:23