Separation and functional analysis of bone marrow cells separated by rhodamine-123 fluorescence

Journal Article · Thu Jan 01 04:00:00 EST 1987 · Exp. Hematol.; (United States)

OSTI ID:7083016

Mulder, A H; Visser, J W

Mouse bone marrow (BM) cells were separated on the basis of fluorescence intensity after labeling with the supravital, fluorescing dye rhodamine 123 (Rh 123). Rh 123 accumulates in the mitochondria. The BM fractions were tested for the presence of spleen colony-forming units (CFU-S) that produced colonies at day 8 and day 12 after bone marrow transplantation; for the ability to rescue lethally irradiated mice; and for thymus-repopulating ability. Results showed that all the day-8 CFU-S incorporated a relatively large amount of Rh 123, while the day-12 CFU-S were stained heterogeneously. Survival after lethal irradiation, as expressed in the numbers of day-12 CFU-S transplanted, was predominately mediated by the weakly fluorescing fraction. However, early thymus repopulation, which is caused by prothymocytes contained in the graft, was mediated by the brightly fluorescing fraction. Data in the literature indicate that a high uptake of RH123 is correlated with cellular proliferation. This suggests that all the day-8 CFU-S and about 60% of the day-12 CFU-S are cycling. In contrast, less than 10% of both day-8 and day-12 CFU-S are killed by S-phase-specific agents. From this we conclude that a high uptake of Rh 123 depends on other factors in addition to the cell cycle status. Differentiation processes in which the day-8 CFU-S appears to be involved also cause the presence of many or very active mitochondria. The difference between the weakly and brightly fluorescing fractions in the number of day-12 CFU-S required for 30-day survival after lethal irradiation, indicate heterogeneity among the day-12 CFU-S population or that the fraction weakly labeled with Rh123 contains other, less mature, cells that are responsible for survival after lethal irradiation and are different from the stem cells that protect lethally irradiated mice.

• Research Organization: Radiobiological Institute TNO, Rijswijk, The Netherlands
• OSTI ID: 7083016
• Journal Information: Exp. Hematol.; (United States), Journal Name: Exp. Hematol.; (United States) Vol. 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.
AMINES
ANIMAL CELLS
ANIMAL TISSUES
ANIMALS
BODY
BONE MARROW
BONE MARROW CELLS
CARBOXYLIC ACIDS
CELL CONSTITUENTS
CFU
CONNECTIVE TISSUE CELLS
DYES
FLUORESCENCE
HEMATOPOIETIC SYSTEM
HETEROCYCLIC ACIDS
HETEROCYCLIC COMPOUNDS
IRRADIATION
LETHAL IRRADIATION
LUMINESCENCE
LYMPHATIC SYSTEM
MAMMALS
MICE
MITOCHONDRIA
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC OXYGEN COMPOUNDS
ORGANOIDS
ORGANS
REAGENTS
RHODAMINES
RODENTS
SOMATIC CELLS
THYMUS
TISSUES
TRANSPLANTS
VERTEBRATES