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Titolo:
Long-term culture of hematopoietic stem cells - validating the stromal component of the CAFC assay
Autore:
Olesen, G; Tonder, H; Holm, MS; Hokland, P;
Indirizzi:
Aarhus Univ Hosp, Dept Hematol, Aarhus, Denmark Aarhus Univ Hosp Aarhus Denmark niv Hosp, Dept Hematol, Aarhus, Denmark
Titolo Testata:
CYTOTHERAPY
fascicolo: 2, volume: 3, anno: 2001,
pagine: 107 - 116
SICI:
1465-3249(2001)3:2<107:LCOHSC>2.0.ZU;2-3
Fonte:
ISI
Lingua:
ENG
Soggetto:
COLONY-STIMULATING FACTOR; LIMITING DILUTION ASSAYS; HUMAN-BONE-MARROW; INITIATING CELLS; FUNCTIONAL-CHARACTERIZATION; PERIPHERAL-BLOOD; HETEROGENEITY; PROGENITORS; LEUKEMIA; SUBSETS;
Keywords:
long-term bone marrow culture; stromal cell source; hematopoietic stem cells; CAFC;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Clinical Medicine
Life Sciences
Citazioni:
35
Recensione:
Indirizzi per estratti:
Indirizzo: Hokland, P Aarhus Kommune Hosp, Immunhaematol Lab, Tage Hansens Gade, Opgang 4A, DK-8000 Aarhus C, Denmark Aarhus Kommune Hosp Opgang 4A Aarhus Denmark C rhus C, Denmark
Citazione:
G. Olesen et al., "Long-term culture of hematopoietic stem cells - validating the stromal component of the CAFC assay", CYTOTHERAPY, 3(2), 2001, pp. 107-116

Abstract

BackgroundThe stroma-based long-term culture is the assay of choice when a functional detection of primitive hematopoietic cells in vitro is sought. However, different stromal cell lines varying in supporting capacity have been raisedand applied in different laboratories, resulting in a wide range in published frequencies of LTCIC alternative CAFC. MethodsIn order to identify the most suitable stromal source in terms of supportive capacity, reproducibility, and ease of handling, we have compared some of the most commonly employed murine cell lines to human bone marrow stroma in secondary long-term culture set-ups. ResultsSeeking an approximation to the supportive capacity of human BM stroma we found the FBMD-1 cell line supplemented with G-CSF and IL-3 superior to FBMD-1 cells alone, and to M2-10B4 and Sl/Sl cells. Moreover, in co-cultures of CD34(+) cells and the FBMD-1 line, we found week 5 CAFC content highly reproducible (50.5+/-6.66-54.6+/-7.07/10(4) plated cells, p value >0.95) and the assay was suitable for inter-individual comparison in a clinical setting. In fact, the week 5 CAFC results were even more reproducible than those of the CFU assays (CV 0.03 for the CAFC assay versus 0.13-0.33 for the CFU assays). On the other hand, when extending the culture period to 8 weeks, the cobblestone area formation was best maintained by human BM stroma and the high reproducibility in CAFC enumeration in cultures supported by the FBMD-1 was lost. DiscussionAmong the stromal cell sources tested, the FBMD-1 line was found to be superior in terms of ease of handling and week 5 CAFC reproducibility. However, this robustness could not be extended to week 8 CAFC.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 24/01/20 alle ore 12:17:47