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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Pluripotent hematopoietic stem cells contain high levels of mRNA for c-kit, GATA-2, p45 NF-E2, and c-myb and low levels or no mRNA for c-fms and the receptors for granulocyte colony-stimulating factor and interleukins 5 and 7.
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Pluripotent hematopoietic stem cells contain high levels of mRNA for c-kit, GATA-2, p45 NF-E2, and c-myb and low levels or no mRNA for c-fms and the receptors for granulocyte colony-stimulating factor and interleukins 5 and 7.

机译：多能造血干细胞含有高水平的c-kit，GATA-2，p45 NF-E2和c-myb mRNA，低水平或无c-fms mRNA，以及粒细胞集落刺激因子和白介素5和7。

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摘要

Pluripotent hematopoietic stem cells (PHSCs) were highly enriched from mouse bone marrow by counterflow centrifugal elutriation, lineage subtraction, and fluorescence-activated cell sorting based on high c-kit receptor expression (c-kitBR). We used reverse transcriptase polymerase chain reaction to assay the c-kitBR subset and the subsets expressing low (c-kitDULL) and no (c-kitNEG) c-kit receptor for expression of mRNA encoding hematopoietic growth factor receptors and transcription factors. The c-kitBR cells had approximately 3.5-fold more c-kit mRNA than unfractionated bone marrow cells. The c-kitDULL cells had 47-58% of the c-kit mRNA found in c-kitBR cells and the c-kitNEG cells had 4-9% of the c-kit mRNA present in c-kitBR cells. By comparing mRNA levels in c-kitBR cells (enriched for PHSCs) with those of unfractionated bone marrow, we demonstrated that c-kitBR cells contained low or undetectable levels of mRNA for c-fms, granulocyte colony-stimulating factor receptor, interleukin 5 receptor (IL-5R), and IL-7R. These same cells had moderate levels of mRNA for erythropoietin receptor, IL-3R subunits IL-3R alpha (SUT-1), AIC-2A, and AIC-2B, IL-6R and its partner gp-130, and the transcription factor GATA-1 and high levels of mRNA for transcription factors GATA-2, p45 NF-E2, and c-myb. We conclude from these findings that PHSCs are programmed to interact with stem cell factor, IL-3, and IL-6 but not with granulocyte or macrophage colony-stimulating factor. These findings also indicate that GATA-2, p45 NF-E2, and c-myb activities may be involved in PHSC maintenance or proliferation.
机译：多能造血干细胞（PHSCs）通过逆流离心淘析，谱系扣除和基于高c-kit受体表达（c-kitBR）的荧光激活细胞分选从小鼠骨髓中高度富集。我们使用逆转录酶聚合酶链反应来检测c-kitBR亚型和表达低（c-kitDULL）和无（c-kitNEG）c-kit受体的亚组，以表达编码造血生长因子受体和转录因子的mRNA。与未分离的骨髓细胞相比，c-kitBR细胞的c-kit mRNA大约多3.5倍。 c-kitDULL细胞的c-kitBR细胞中的c-kit mRNA为47-58％，c-kitNEG细胞的c-kitBR细胞中的c-kit mRNA为4-9％。通过比较c-kitBR细胞（富含PHSC）和未分离的骨髓中的mRNA水平，我们证明了c-kitBR细胞中c-fms，粒细胞集落刺激因子受体，白介素5受体的mRNA水平较低或无法检测。 （IL-5R）和IL-7R。这些相同细胞的促红细胞生成素受体，IL-3R亚基IL-3R alpha（SUT-1），AIC-2A和AIC-2B，IL-6R及其伴侣gp-130和转录因子GATA的mRNA水平中等-1和转录因子GATA-2，p45 NF-E2和c-myb的高水平mRNA。我们从这些发现得出结论，PHSCs被编程为与干细胞因子，IL-3和IL-6相互作用，但不与粒细胞或巨噬细胞集落刺激因子相互作用。这些发现还表明，GATA-2，p45 NF-E2和c-myb活性可能与PHSC的维持或增殖有关。

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