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Bulk enrichment of transplantable hemopoietic stem cell subsets from lipopolysaccharide-stimulated murine spleen

Journal Article · · Exp. Hematol.; (United States)
OSTI ID:6858057
; ;
Counterflow centrifugal elutriation (CCE) in combination with density flotation centrifugation and fluorescence-activated cell sorting on wheat-germ agglutinin-FITC(WGA)-binding cells within the light-scatter ''blast window'' were used consecutively to enrich pluripotent hemopoietic stem cells (HSC) in bulk from lipopolysaccharide-stimulated mouse spleen. The medium-to-strong WGA + ve fraction contained 3.10(6) cells isolated from 3-4 X 10(9) spleen cells, with an average of 126% day-12 CFU-S and 65% day-8 CFU-S as calculated on the basis of their seeding fraction, suggesting that virtually all cells represented in vivo macroscopic colony formers. In view of the large differences reported elsewhere between stem cell subsets differing in reconstitutive capacity and secondary stem cell generation ability, we also studied various isolated cell fractions with respect to spleen colony formation, radioprotective ability, and spleen- and marrow- repopulating ability. Day-8 and day-12 CFU-S copurified when isolated by CCE. Cells from a fraction with high affinity for WGA were most highly enriched for their radioprotective ability (RPA) and their ability to repopulate the cellularity of the spleen and femur of irradiated recipients. This fraction contained virtually pure day-12 CFU-S. However, the ability to generate secondary day-12 CFU-S and CFU-GM in irradiated organs was enriched most in the medium WGA + ve cell fraction. MRA and SRA, according to the latter criteria, could therefore be partly separated from day-12 CFU-S and RPA on the basis of affinity for WGA. The data strongly suggest that at least part of all day-12 CFU-S have a high potential to proliferate and differentiate into mature progeny, but a relatively low self-renewal ability, and may therefore not be representative of the genuine stem cell.
Research Organization:
Erasmus Univ., Rotterdam, The Netherlands
OSTI ID:
6858057
Journal Information:
Exp. Hematol.; (United States), Journal Name: Exp. Hematol.; (United States) Vol. 2; ISSN EXHMA
Country of Publication:
United States
Language:
English

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Related Subjects

560152* -- Radiation Effects on Animals-- Animals
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ANIMAL CELLS
ANIMALS
CARBOHYDRATES
CELL DIVISION
CELL FLOW SYSTEMS
CELL PROLIFERATION
CENTRIFUGATION
COLONY FORMATION
FLUORESCENCE
IRRADIATION
LIPIDS
LIPOPOLYSACCHARIDES
LUMINESCENCE
MAMMALS
MICE
ORGANIC COMPOUNDS
POLYSACCHARIDES
RODENTS
SACCHARIDES
SEPARATION PROCESSES
SOMATIC CELLS
SPLEEN CELLS
STEM CELLS
TRANSPLANTS
VERTEBRATES