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a Institute of Human Genetics, University of Newcastle upon Tyne, International Centre for Life, Newcastle upon Tyne, United Kingdom;
b BD Biosciences, Oxford, United Kingdom;
c Department of Biological Sciences, University of Durham, Durham, United Kingdom
Key Words. Hematopoietic stem cells • Aldehyde dehydrogenase activity • Telomerase activity • Murine hematopoietic progenitor cells
Correspondence: Majlinda Lako, Ph.D., Institute of Human Genetics, University of Newcastle upon Tyne, International Centre for Life, Central Parkway, Newcastle upon Tyne NE1 3BZ, U.K. Telephone: 00-44-191-241-8688; Fax: 00-44-191-241-8666; e-mail: Majlinda.Lako{at}ncl.ac.uk
There are several different technical approaches to the isolation of hematopoietic stem cells (HSCs) with long-term repopulating ability, but these have problems in terms of yield, complexity, or cell viability. Simpler strategies for HSC isolation are needed. We have enriched primitive hematopoietic progenitors from murine bone marrow of mice from different genetic backgrounds by lineage depletion followed by selection of cells with high aldehyde dehydrogenase activity using the Aldefluor reagent (BD Biosciences, Oxford, U.K.). Lin–ALDHbright cells comprised 26.8 ± 1.0% of the total Lin– population of C57BL6 mice, and 23.5 ± 1.0% of the Lin– population of BALB/c mice expressed certain cell-surface markers typical of primitive hematopoietic progenitors. In vitro hematopoietic progenitor function was substantially higher in the Lin–ALDHbright population compared with the Lin–ALDHlow cells. These cells have higher telomerase activity and the lowest percentage of cells in S phase. These data strongly suggest that progenitor enrichment from Lin–cells on the basis of ALDH is a valid method whose simplicity of application makes it advantageous over conventional separations.
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