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Further Characterization of CD34‐Low/Negative Mouse Hematopoietic Stem Cells
Author(s) -
NAKAUCHI HIROMITSU,
TAKANO HINA,
EMA HIDEO,
OSAWA MASATAKE
Publication year - 1999
Publication title -
annals of the new york academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.712
H-Index - 248
eISSN - 1749-6632
pISSN - 0077-8923
DOI - 10.1111/j.1749-6632.1999.tb08453.x
Subject(s) - cd34 , haematopoiesis , stem cell , biology , microbiology and biotechnology , stem cell factor , hematopoietic stem cell , progenitor cell , bone marrow , cell culture , chemistry , immunology , genetics
A bstract : We have previously reported that in adult mouse bone marrow, CD34 low/− c‐kit + Sca‐1 + lineage markers negative (Lin − ) (CD34 − KSL) cells represent hematopoietic stem cells with long‐term marrow repopulating ability whereas CD34 + c‐kit + Sca‐1 + Lin − (CD34 + KSL) cells are progenitors with short‐term reconstitution capacity. To further characterize cells in those two populations, relative expression of various genes wereee examniend by reverse transcriptase polymerase chain reaction (RT‐PCR). In CD34 − KSL Cells, none of the genes sturied was found to be expressed with the exception of GATA‐2, IL‐1Rα, IL‐2Rγ, AIC‐2B, c‐kit, EPO‐R, and c‐mpl. In contrast, expression of GATA‐1 and all cytokine receptor genes examined except IL‐2Rβ, IL‐7Rα and IL‐9Rα were found in CD34 + KSL. The difference between these two populations was also shown in single cell culture analysis of these cells. when cells were clone sorted and cultured in the presence of SCF, IL‐3 and EPO, CD34 − KSL cells required much more time to undergo the first cell division than CD34 + KSL cells. dormancy and random fashion of cell division by CD34 − KSL cells were also evident by the analysis of the second cell division, which was found to be delayed and unsynchronous comapred with CD34 + KSL cells. Clonal culture analusis showed that CD34 − KSL cells were more potent in proliferation and multilineage differentiation capacities than CD34 + KSL cells. In a paired‐daughter cell experiment, 75% of CD34 − KSL and 50% of CD34 + KSL paired‐daughter‐derived colonies were nonidentical wiht wide vairety of lineage combinations. Taken together, these data support our previous notion that CD34 − KSL cells are at higher rank in hematopoietic hierarchy than CD34 + KSL cells. In addition, our results using highly enriched stem cell population directly obtained from mouse bone marrow support the proposed stochastic nature of lineage commitment.