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Titolo:
QUANTITATION OF THE QUIESCENT FRACTION OF LONG-TERM CULTURE-INITIATING CELLS IN NORMAL HUMAN BLOOD AND MARROW AND THE KINETICS OF THEIR GROWTH FACTOR-STIMULATED ENTRY INTO S-PHASE IN-VITRO
Autore:
PONCHIO L; CONNEALLY E; EAVES C;
Indirizzi:
BRITISH COLUMBIA CANC AGCY,TERRY FOX LAB,601 W 10TH AVE VANCOUVER BC V5Z 1L3 CANADA BRITISH COLUMBIA CANC AGCY,TERRY FOX LAB VANCOUVER BC V5Z 1L3 CANADA UNIV BRITISH COLUMBIA,DEPT MED GENET VANCOUVER BC CANADA UNIV BRITISH COLUMBIA,DEPT PATHOL & LAB MED VANCOUVER BC V5Z 1M9 CANADA
Titolo Testata:
Blood
fascicolo: 9, volume: 86, anno: 1995,
pagine: 3314 - 3321
SICI:
0006-4971(1995)86:9<3314:QOTQFO>2.0.ZU;2-F
Fonte:
ISI
Lingua:
ENG
Soggetto:
HEMATOPOIETIC STEM-CELLS; LIMITING-DILUTION; PROGENITOR CELLS; CFU-S; PROLIFERATION; SEPARATION; INVITRO; CYCLE; LINE;
Tipo documento:
Note
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
31
Recensione:
Indirizzi per estratti:
Citazione:
L. Ponchio et al., "QUANTITATION OF THE QUIESCENT FRACTION OF LONG-TERM CULTURE-INITIATING CELLS IN NORMAL HUMAN BLOOD AND MARROW AND THE KINETICS OF THEIR GROWTH FACTOR-STIMULATED ENTRY INTO S-PHASE IN-VITRO", Blood, 86(9), 1995, pp. 3314-3321

Abstract

A method for quantitating the proportion of cycling longterm culture-initiating cells (LTC-IC) in heterogeneous populations of human hematopoietic cells is described. This procedure involves incubating the cells of interest for 16 to 24 hours in a serum-free medium containing 100 ng/mL Steel factor (SF), 20 ng/mL interleukin-3 (IL-3), and 20 ng/mLgranulocyte-colony-stimulating factor (G-CSF), with or without 20 mu Ci/mL of high specific activity H-3-thymidine (H-3-Tdr) before platingthe recovered cells in standard LTC-IC assays. The details of this procedure are based in part on the finding that the number of LTC-IC (regardless of their cycling status) remains constant for at least 24 hours under these culture conditions, as long as H-3-Tdr is not present. In addition, we have determined that a 16-hour period of exposure to the H-3-Tdr is sufficient to maximize the discrimination of cycling LTC-IC but not long enough to allow a detectable redistribution of LTC-ICbetween noncycling and cycling compartments. Finally, any isotope reutilization that may occur is not sufficient to affect the LTC-IC H-3-Tdr suicide values measured. Application of this methodology to normally circulating LTC-IC showed these to be a primarily quiescent population. However, within 72 hours of incubation in a serum-free medium containing SF, IL-3, and G-CSF, most had entered S-phase, although there was no net change in their numbers. This suggests that, under certain conditions in vitro, self-renewal divisions of LTC-IC can occur and, atleast initially, balance any losses of these cells due to their differentiation or death. In contrast, many of the LTC-IC in freshly aspirated samples of normal marrow were found to be proliferating, although those that were initially quiescent could also be recruited into S-phase within 72 hours in vitro when incubated under the same conditions used to stimulate circulating LTC-IC. This modified H-3-Tdr suicide procedure should facilitate further investigation of the mechanisms regulating the turnover of the most primitive compartments of human hematopoietic cells and how these may be altered in disease states or exploited for a variety of therapeutic applications. (C) 1995 by The AmericanSociety of Hematology.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 27/01/20 alle ore 01:18:01