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Titolo:
EFFICIENT ISOLATION OF HUMAN CD34 POSITIVE HEMATOPOIETIC PROGENITOR CELLS BY IMMUNE PANNING
Autore:
HOLYOAKE TL; ALCORN MJ; RICHMOND LJ; FRESHNEY MG; PEARSON C; FITZSIMONS E; STEWARD WP; DUNLOP DJ; PRAGNELL IB;
Indirizzi:
BEATSON CRC LABS,GARSCUBE ESTATE,SWITCHBACK RD GLASGOW G61 1BD SCOTLAND BEATSON INST CANC RES GLASGOW SCOTLAND GLASGOW ROYAL INFIRM,LEUKAEMIA RES FUND LABS GLASGOW SCOTLAND MONKLANDS DIST GEN HOSP,DEPT HAEMATOL AIRDRIE SCOTLAND WESTERN INFIRM & ASSOCIATED HOSP,DEPT MED ONCOL GLASGOW SCOTLAND
Titolo Testata:
Stem cells
fascicolo: 1, volume: 12, anno: 1994,
pagine: 114 - 124
SICI:
1066-5099(1994)12:1<114:EIOHCP>2.0.ZU;2-T
Fonte:
ISI
Lingua:
ENG
Soggetto:
BONE-MARROW TRANSPLANTATION; HEMATOPOIETIC STEM-CELLS; GROWTH-FACTOR; CFU-GM; PROLIFERATION; INVITRO; ENGRAFTMENT; CANCER; ASSAY;
Keywords:
CD34; IMMUNE PANNING; STEM CELLS; CLONOGENIC ASSAYS; BONE MARROW TRANSPLANTATION; BFU-E; CFU-GM; CFU-A;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Science Citation Index Expanded
Citazioni:
26
Recensione:
Indirizzi per estratti:
Citazione:
T.L. Holyoake et al., "EFFICIENT ISOLATION OF HUMAN CD34 POSITIVE HEMATOPOIETIC PROGENITOR CELLS BY IMMUNE PANNING", Stem cells, 12(1), 1994, pp. 114-124

Abstract

In this study we have assessed the use of soybean agglutinin (SBA) and CD34 microcellector devices for the selection of CD34 positive hemopoietic progenitor cells. Burst forming unit-erythroid (BFU-E), colony forming unit-granulecyte/macrophage (CFU-GM) and the recently developed multipotential human colony forming unit-type A (CFU-A) clonogenic assays were used to measure progenitor numbers in the starting mononuclear cell (MNC), the SBA negative, the nonadherent CD34 negative and the adherent CD34 positive fractions during panning. CFU-A progenitors were present at a relatively high incidence in the MNC fraction (220 per 10(5) MNC) and were enriched 15-fold in the adherent CD34 positive fraction. This progenitor incidence and enrichment were similar to those of CFU-GM and BFU-E. The mean recovery for CD34 positive cells was 2.3 x 10(6) cells per marrow aspirate. Analyses by flow cytometry demonstrated that 1-5% of input MNC were CD34 positive, that the purity of the CD34 fraction was approximately 80% and that the calculated recovery for CD34 positive cells was 61%. Recoveries for CFU-GM, BFU-E and CFU-A were between 18 and 40%. CFU-A progenitors were found exclusivelyin the adherent CD34 positive fraction, whereas a significant proportion of both CFU-GM and BFU-E were present in the nonadherent CD34 negative fraction. We propose that the Applied Immune Sciences (AIS) flasks preferentially bind the cells which express CD34 most strongly and that this is reflected in the finding of primitive CFU-A only in the CD34 positive fraction, with lineage-restricted progenitors found in both CD34 positive and negative fractions. This hypothesis is strengthened by data on long-term bone marrow cultures in which the CD34 positivefraction is better able to maintain output of CFU-GM compared with the CD34 negative fraction. In conclusion, relatively pure populations of CD34 positive cells may be rapidly and efficiently isolated from bone marrow samples with good recovery. The isolated cells show enhanced colony forming capacity in standard clonogenic assays and in the multipotential CFU-A assay.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 18/09/20 alle ore 17:11:51