Long-term monoclonal reconstitution of erythropoiesis in genetically anemic W/Wv mice by injection of 5-fluorouracil-treated bone marrow cells of Pgk-1b/Pgk-1a mice
The spleen colony-forming assay does not represent the number of hematopoietic stem cells with extensive self-maintaining capacity because five to 50 spleen colony-forming units (CFU-S) are necessary to rescue a genetically anemic (WB X C57BL/6)F1-W/Wv(WBB6F1-W/Wv) mouse. We investigated which is more important for the reconstitution of erythropoiesis, the transplantation of multiple CFU-S or that of a single stem cell with extensive self-maintaining potential. The electrophoretic pattern of hemoglobin was used as a marker of reconstitution and that of phosphoglycerate kinase (PGK), an X chromosome-linked enzyme, as a tool for estimating the number of stem cells. For this purpose, we developed the C57BL/6 congeneic strain with the Pgk-1a gene. Bone marrow cells were harvested after injection of 5-fluorouracil from C57BL/6-Pgk-1b/Pgk-1a female mice in which each stem cell had either A-type PGK or B-type PGK due to the random inactivation of one or two X chromosomes. When a relatively small number of bone marrow cells (ie, 10(3) or 3 X 10(3) were injected into 200-rad-irradiated WBB6F1-W/Wv mice, the hemoglobin pattern changed from the recipient type (Hbbd/Hbbs) to the donor type (Hbbs/Hbbs) in seven of 150 mice for at least 8 weeks. Erythrocytes of all these WBB6F1-W/Wv mice showed either A-type PGK alone or B-type PGK alone during the time of reconstitution, which suggests that a single stem cell with extensive self-maintaining potential may sustain the whole erythropoiesis of a mouse for at least 8 weeks.
- Research Organization:
- Osaka Univ. Medical School, Japan
- OSTI ID:
- 5517416
- Journal Information:
- Blood; (United States), Vol. 70:6
- Country of Publication:
- United States
- Language:
- English
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BONE MARROW CELLS
TRANSPLANTS
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DYNAMIC FUNCTION STUDIES
ANEMIAS
COLONY FORMATION
ERYTHROCYTES
FLUOROURACILS
HEMOGLOBIN
MICE
PHOSPHOTRANSFERASES
RADIATION CHIMERAS
STEM CELLS
X CHROMOSOME
ANIMAL CELLS
ANIMALS
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BLOOD
BLOOD CELLS
BLOOD FORMATION
BODY FLUIDS
CARBOXYLIC ACIDS
CHIMERAS
CHROMOSOMES
CONNECTIVE TISSUE CELLS
DISEASES
DRUGS
ENZYMES
GLOBIN
HEMIC DISEASES
HETEROCHROMOSOMES
HETEROCYCLIC ACIDS
HETEROCYCLIC COMPOUNDS
HYDROXY COMPOUNDS
MAMMALS
MATERIALS
MOSAICISM
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC FLUORINE COMPOUNDS
ORGANIC HALOGEN COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PHOSPHORUS-GROUP TRANSFERASES
PIGMENTS
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SYMPTOMS
TRANSFERASES
URACILS
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560152* - Radiation Effects on Animals- Animals