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Expression of hyaluronan synthase genes in umbilical cord blood stem/progenitor cells

机译:透明质酸合酶基因在脐血干/祖细胞中的表达

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Scientific progress reveals an ever-expanding role of hyaluronan (HA) in diverse biological functions. It has become increasingly clear that HA might also be essential for certain functions of stem cells. CD133+ cells isolated from umbilical cord blood (UCB) seem to represent an alternative to CD34+ cells as a source of transplantable haematopoietic progenitor cells. The aim of this study was to investigate expression patterns of hyaluronan synthases (HAS) genes in freshly isolated and cultured UCB progenitor cells and to compare HAS mRNA levels to those found in non-progenitor cells. CD133+ stem cells were isolated from UCB using an immumomagnetic procedure. Investigation of HAS mRNA expression patterns in CD 133+ and CD 133- cells by RT-PCR was performed immediately after isolation as well as after cultivation towards myelomonocytic lineage. In addition, activation patterns of mitogen activated protein kinases (MAPK) were analyzed by Western blot experiments. mRNA for HAS1 is undetectable but HAS3 mRNA can be readily detected in freshly isolated CD133+ as well as in CD133- UCB cells. More importantly, our data demonstrate that mRNA for HAS2 can only be detected in CD 133+ progenitor cells. In addition, while MAPK are slightly activated in CD133- UCB cells, no significant phosphorylation of MAPK could be observed in CD133+ cells, excluding a role of these kinases in the regulation of HAS2. HAS2 is expressed only in freshly isolated CD133+ cells and quickly diminishes during differentiation. Because of this, HAS2 gene expression might be suitable as a new marker for CD133+ UCB-derived stem cells. (c) 2006 Elsevier B.V. All rights reserved.
机译:科学进步揭示了透明质酸(HA)在多种生物学功能中的不断扩大的作用。越来越清楚的是,HA对于干细胞的某些功能可能也必不可少。从脐带血(UCB)分离出的CD133 +细胞似乎代表着CD34 +细胞的替代品,作为可移植造血祖细胞的来源。这项研究的目的是调查透明质酸合酶(HAS)基因在新鲜分离和培养的UCB祖细胞中的表达模式,并将HAS mRNA水平与非祖细胞中发现的水平进行比较。使用免疫磁方法从UCB分离CD133 +干细胞。在分离后以及向骨髓单核细胞系培养后立即通过RT-PCR研究CD 133+和CD 133-细胞中HAS mRNA表达模式。另外,通过蛋白质印迹实验分析了促分裂原活化蛋白激酶(MAPK)的活化模式。无法检测到HAS1的mRNA,但可以在新鲜分离的CD133 +和CD133-UCB细胞中轻松检测到HAS3 mRNA。更重要的是,我们的数据表明,仅在CD 133+祖细胞中可以检测到HAS2的mRNA。另外,尽管MAPK在CD133-UCB细胞中被轻微激活,但是在CD133 +细胞中未观察到MAPK的显着磷酸化,除了这些激酶在HAS2调节中的作用之外。 HAS2仅在新鲜分离的CD133 +细胞中表达,并在分化过程中迅速减少。因此,HAS2基因表达可能适合作为CD133 + UCB衍生干细胞的新标记。 (c)2006 Elsevier B.V.保留所有权利。

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