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Titolo:
PRIMITIVE HUMAN HEMATOPOIETIC-CELLS DISPLAYING DIFFERENTIAL EFFLUX OFTHE RHODAMINE-123 DYE HAVE DISTINCT BIOLOGICAL-ACTIVITIES
Autore:
UCHIDA N; COMBS J; CHEN S; ZANJANI E; HOFFMAN R; TSUKAMOTO A;
Indirizzi:
SYSTEMIX INC,3155 PORTER DR PALO ALTO CA 94304 UNIV NEVADA,DEPT VET AFFAIRS RENO NV 89557
Titolo Testata:
Blood
fascicolo: 4, volume: 88, anno: 1996,
pagine: 1297 - 1305
SICI:
0006-4971(1996)88:4<1297:PHHDDE>2.0.ZU;2-Y
Fonte:
ISI
Lingua:
ENG
Soggetto:
STEM-CELLS; BONE-MARROW; MULTIDRUG RESISTANCE; PROGENITOR CELLS; SEPARATION; EXPRESSION; SPLEEN; SHEEP; MICE; LIN;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
27
Recensione:
Indirizzi per estratti:
Citazione:
N. Uchida et al., "PRIMITIVE HUMAN HEMATOPOIETIC-CELLS DISPLAYING DIFFERENTIAL EFFLUX OFTHE RHODAMINE-123 DYE HAVE DISTINCT BIOLOGICAL-ACTIVITIES", Blood, 88(4), 1996, pp. 1297-1305

Abstract

Human bone marrow (BM) CD34(+) cells were stained with the vital dye,rhodamine 123 (Rh123), and analyzed for their biological properties based on the level of dye retention. Heterogeneous rhodamine staining is seen within the CD34(+) population, and the staining patterns differdramatically between fetal BM (FBM), adult BM (ABM) and mobilized peripheral blood (MPB). Kinetic analysis of the efflux of Rh123 from ABM CD34(+) cells showed that efflux of Rh123 was most rapid from the mostprimitive Thy-1(+) subset. The efflux of Rh123 could be inhibited by verapamil, suggesting that rhodamine efflux from primitive hematopoietic cells is primarily due to the P-glycoprotein (P-gp) pump or anotherintracellular transport system affected by verapamil. When four CD34() subpopulations were plated onto SyS1 BM stromal cell cocultures after 1 to 2 weeks, only wells plated with CD34(+)Thy-1(+)Rh123(lo) (low-level Rh123 retention) or CD34(+)Thy-1(+)Rh123(mid) (mid-level Rh123 retention) cells maintained greater than 50% of cells in an uncommittedCD34(+)33(-) stage. CD34(+)Lin(-) (lineage-negative) cells were fractionated based on Rh123 dye staining into Rh123(hi) (high-level Rh123 retention), Rh123(mid), and Rh123(lo) and deposited as single cells into long-term SyS1 BM stromal cell cultures. The Rh123(mid) fraction hadimmense early proliferative activity in vitro, but lost the ability to form cobblestone areas after 5 to 6 weeks in culture. In contrast, the Rh123(lo) fraction proliferated more slowly but sustained long-termin vitro hematopoiesis as evidenced by continued cobblestone area-forming cells (CAFC) activity for at least 6 weeks. The Rh123(hi) fraction showed a plating efficiency similar to that of the Rh123(lo) or Rh123(mid) fractions but did not extensively proliferate in vitro and did not show evidence of CAFC activity, We predicted from these in vitro results that the Rh123(lo) subset possesses long-term engrafting potential. Indeed, on transplantation into the SCID-hu bone assay, all long-term engrafting potential and multilineage differentiation potential resided within the Rh1231(lo-mid) but not Rh123(hi) subset. Furthermore, human marrow subpopulations derived from chimeric sheep after in utero transplantation with CD34(+)Thy-1(+)Lin(-) cells were reisolated based on Rh123 staining. Again, CD34(+)Lin(-) subsets showing Rh123(lo-mid) had long-term growth in culture, whereas Rh123(hi)CD34(+)Lin(-) cells did not. These results show that, after injection of CD34(+)Thy-1()Lin(-) cells into an in utero microenvironment, primitive CD34(+) cells maintain a Rh123 phenotype that correlates with their in vitro CAFC activity. Thus, Rh123 staining is an effective way to define functional subsets of primitive hematopoietic cell populations. (C) 1996 by The American Society of Hematology.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 14/07/20 alle ore 10:11:30