Isolation of hemopoietic stem cell subsets from murine bone marrow: II. Evidence for an early precursor of day-12 CFU-S and cells associated with radioprotective ability

Journal Article · Fri Jan 01 04:00:00 EST 1988 · Exp. Hematol.; (United States)

OSTI ID:5592068

Ploemacher, R E; Brons, N H

Counterflow centrifugal elutriation (CCE) in combination with plastic adherence and fluorescence-activated cell sorting were used consecutively to enrich functionally different subpopulations of pluripotent hemopoietic stem cells (HSC) from mouse bone marrow. The nonadherent CCE fractions were labeled with wheat germ agglutinin (WGA)-fluorescein isothiocyanate (FITC) and sorted according to differences in fluorescence within various windows on the basis of forward (FLS) and perpendicular (PLS) light scatter. The sorted cells were then assayed for their (1) in vivo colony-forming ability (day-7 and day-12 spleen colony-forming units (CFU-S)), (2) radioprotective ability (RPA; 30-day survival), and (3) their ability to repopulate the bone marrow or spleen over a 13-day period with day-12 CFU-S, granulocyte-macrophage colony-forming units (CFU-GM), nucleated cells, or cells associated with RPA. The highest incidence of day-12 CFU-S and cells with RPA was obtained by sorting the most WGA-positive cells with relatively high PLS (enrichment, 50- to 200-fold), lowering the effective dose (ED 50/30) to an average of 80 cells. The separative procedure enabled hemopoietic stem cells that repopulate both bone marrow and spleen with secondary RPA cells, CFU-S-12, and CFU-GM to be enriched and separated from part of the RPA cells, CFU-S-12, and cells that reconstitute the cellularity of bone marrow and spleen. These data suggest that cells generating both day-12 CFU-S and RPA cells differ from day-12 CFU-S and RPA cells themselves on the basis of PLS characteristics and affinity for WGA. Such early stem cells have also been detected in sorted fractions meeting the FLS/PLS characteristics of lymphocytes.

• Research Organization: Erasmus Univ., Rotterdam, The Netherlands
• OSTI ID: 5592068
• Journal Information: Exp. Hematol.; (United States), Journal Name: Exp. Hematol.; (United States) Vol. 16:1; ISSN EXHMA
• Country of Publication: United States
• Language: English

Related Subjects

560152* -- Radiation Effects on Animals-- Animals
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
AGGLUTININS
ANIMAL CELLS
ANIMAL TISSUES
ANIMALS
ANTIBODIES
BODY
BONE MARROW
CENTRIFUGATION
COLONY FORMATION
HEMATOPOIETIC SYSTEM
MAMMALS
MICE
ORGANS
PRECURSOR
RADIATION PROTECTION
RADIOSENSITIVITY EFFECTS
RODENTS
SCATTERING
SEPARATION PROCESSES
SOMATIC CELLS
SPLEEN
STEM CELLS
TISSUES
VERTEBRATES