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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
3
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pubmed:dateCreated |
1989-9-7
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pubmed:abstractText |
A simple procedure is described for the quantitation and enrichment of murine hematopoietic cells with the capacity for long-term repopulation of lymphoid and myeloid tissues in lethally irradiated mice. To ensure detection of the most primitive marrow cells with this potential, we used a competitive assay in which female recipients were injected with male "test" cells and 1 to 2 x 10(5) "compromised" female marrow cells with normal short-term repopulating ability, but whose long-term repopulating ability had been reduced by serial transplantation. Primitive hematopoietic cells were purified by flow cytometry and sorting based on their forward and orthogonal light-scattering properties, and Thy-1 and H-2K antigen expression. Enrichment profiles for normal marrow, and marrow of mice injected with 5-fluorouracil (5-FU) four days previously, were established for each of these parameters using an in vitro assay for high proliferative potential, pluripotent colony-forming cells. When all four parameters were gated simultaneously, these clonogenic cells were enriched 100-fold. Both day 9 and day 12 CFU-S were copurified; however, the purity (23%) and enrichment (75-fold) of day 12 CFU-S in the sorted population was greater with 5-FU-treated cells. Five hundred of the sorted 5-FU marrow cells consistently repopulated recipient lymphoid and myeloid tissues (greater than 50% male, 1 to 3 months post-transplant) when co-injected with 1 to 2 x 10(5) compromised female marrow cells, and approximately 100 were sufficient to achieve the same result in 50% of recipients under the same conditions. This relatively simple purification and assay strategy should facilitate further analysis of the heterogeneity and regulation of stem cells that maintain hematopoiesis in vivo.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
AIM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Aug
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pubmed:issn |
0006-4971
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
15
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pubmed:volume |
74
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
930-9
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pubmed:dateRevised |
2006-11-15
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pubmed:meshHeading |
pubmed-meshheading:2568865-Animals,
pubmed-meshheading:2568865-Antigens, Surface,
pubmed-meshheading:2568865-Antigens, Thy-1,
pubmed-meshheading:2568865-Cell Separation,
pubmed-meshheading:2568865-Cells, Cultured,
pubmed-meshheading:2568865-Colony-Forming Units Assay,
pubmed-meshheading:2568865-Female,
pubmed-meshheading:2568865-Flow Cytometry,
pubmed-meshheading:2568865-H-2 Antigens,
pubmed-meshheading:2568865-Hematopoiesis,
pubmed-meshheading:2568865-Hematopoietic Stem Cells,
pubmed-meshheading:2568865-Light,
pubmed-meshheading:2568865-Male,
pubmed-meshheading:2568865-Mice,
pubmed-meshheading:2568865-Phenotype,
pubmed-meshheading:2568865-Scattering, Radiation
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pubmed:year |
1989
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pubmed:articleTitle |
Isolation in a single step of a highly enriched murine hematopoietic stem cell population with competitive long-term repopulating ability.
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pubmed:affiliation |
Terry Fox Laboratory, B. C. Cancer Research Centre, Vancouver.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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