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Titolo:
Cell separation improves the sensitivity of detecting rare human normal and leukemic hematopoietic cells in vivo in NOD/SCID mice
Autore:
Holyoake, TL; Horrocks, C; Thomas, T; Eaves, CJ; Eaves, AC;
Indirizzi:
Royal Infirm, Dept Med, ATMU, Glasgow G31 2ER, Lanark, Scotland Royal Infirm Glasgow Lanark Scotland G31 2ER ow G31 2ER, Lanark, Scotland StemCell Technol Inc, Vancouver, BC, Canada StemCell Technol Inc Vancouver BC Canada hnol Inc, Vancouver, BC, Canada Univ British Columbia, Dept Med Genet, British Columbia Canc Agcy, Terry Fox Lab, Vancouver, BC V5Z 1M9, Canada Univ British Columbia Vancouver BC Canada V5Z 1M9 ver, BC V5Z 1M9, Canada Univ British Columbia, Dept Med, British Columbia Canc Agcy, Terry Fox Lab, Vancouver, BC V5Z 1M9, Canada Univ British Columbia Vancouver BC Canada V5Z 1M9 ver, BC V5Z 1M9, Canada
Titolo Testata:
CYTOTHERAPY
fascicolo: 6, volume: 2, anno: 2000,
pagine: 411 - 421
SICI:
1465-3249(2000)2:6<411:CSITSO>2.0.ZU;2-7
Fonte:
ISI
Lingua:
ENG
Soggetto:
CHRONIC MYELOID-LEUKEMIA; HUMAN CORD-BLOOD; COMBINED IMMUNODEFICIENCY MICE; COLONY-STIMULATING FACTOR; STEM-CELLS; CHRONIC-PHASE; BONE-MARROW; PROGENITOR CELLS; PERIPHERAL-BLOOD; CD34(+) CELLS;
Keywords:
stem cells; cell separation; NOD/SCID; xenotransplants;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Clinical Medicine
Life Sciences
Citazioni:
34
Recensione:
Indirizzi per estratti:
Indirizzo: Holyoake, TL Royal Infirm, Dept Med, ATMU, 10 Alexandra Parade, Glasgow G31 2ER, Lanark, Scotland Royal Infirm 10 Alexandra Parade Glasgow Lanark Scotland G31 2ER
Citazione:
T.L. Holyoake et al., "Cell separation improves the sensitivity of detecting rare human normal and leukemic hematopoietic cells in vivo in NOD/SCID mice", CYTOTHERAPY, 2(6), 2000, pp. 411-421

Abstract

BackgroundThis report describes a novel cell-separation procedure developed to improve detection and analysis of rare human hematopoietic populations, obtainedfrom NOD/SCID mice engrafted with normal and/or leukemic stem cells. MethodsIn preliminary experiments, artificial mixtures of murine and human BM cells were labeled with a combination of Abs specific for murine hematopoieticcells, prior to immunomagnetic negative selection using StemSep. In subsequent experiments, BM was harvested from individual NOD/SCID mice transplanted 6-12 weeks earlier with either human cord blood or primary CML cells anda similar immunomagnetic selection procedure was applied to enrich human cells present. ResultsApplication of this selection procedure to mixtures of murine and human hematopoietic cells using anti-mouse CD45 and Ter-119 allowed a > 1000-fold depletion of murine cells with > 50% recovery of human cells, including progenitors. This level of depletion and recovery were found to be reproduciblefor NOD/SCID mice transplanted and engrafted with human cord blood stem cells, thus facilitating detection of human progenitors, including colony-forming cells (CFC) and LTCIC. For NOD/SCID mice previously transplanted with CML cells, this procedure increased the sensitivity of detective rare humancell subsets by up to > 100-fold. This, in turn, improved the sensitivity of RT-PCR for BCR-ABL and made possible the identification by FACS of various minor subsets of human cells, including CD34(-)CD19/20(+) B-lineage cells, CD34(+) progenitors, mature CD15(+) myeloid cells and CD3(+) T cells present in the mice. DiscussionThis simple cell-depletion procedure should facilitate future investigations of normal and CML stem cell populations in vitro and in NOD/SCID mice.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 15/08/20 alle ore 19:47:56