Further characterization of CD34‐low/negative mouse hematopoietic stem cells

H Nakauchi, H Takano, H Ema… - Annals of the New York …, 1999 - Wiley Online Library
H Nakauchi, H Takano, H Ema, M Osawa
Annals of the New York Academy of Sciences, 1999Wiley Online Library
We have previously reported that in adult mouse bone marrow, CD34low/− 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 …
Abstract: We have previously reported that in adult mouse bone marrow, CD34low/− c‐kit+ Sca‐1+ lineage markers negative (Lin) (CD34KSL) 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 CD34KSL 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, CD34KSL cells required much more time to undergo the first cell division than CD34+KSL cells. dormancy and random fashion of cell division by CD34KSL 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 CD34KSL cells were more potent in proliferation and multilineage differentiation capacities than CD34+KSL cells. In a paired‐daughter cell experiment, 75% of CD34KSL 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 CD34KSL 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.
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