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Analysis of the maintenance of hematopoietic stem cells during embryogenesis

胚胎发生过程中造血干细胞的维持分析

基本信息

批准号：
22790915
负责人：
SUGIYAMA Daisuke
金额：
$ 2.58万
依托单位：
Kyushu University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Young Scientists (B)
财政年份：
2010
资助国家：
日本
起止时间：
2010 至 2011
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-22790915/
关键词：
血液内科学造血幹細胞造血発生血液発生肝臓分化

项目摘要

1. Analysis of KS-13 signal. By database of KS-13, we identified Hmgn2, which gene expression is down-regulated during erythroid differentiation. Over-expression of Hmgn2 gene into leukemic cell lines resulted in suppression of erythroid differentiation in vitro(Kulkeaw et al., 2012).2. Identification of novel molecules that regulate differentiation of hematopoietic stem cells. Fetal liver comprises hematopoietic cells, hepatoblasts and sinusoid endothelial cells. By Flow cytometry, we sorted out Dlk-1(+) hepatoblasts, which primarily expressed extra-cellular matrix such as fibronectin and vitronectin, in addition to hematopoietic cytokines, SCF and EPO(Sugiyama et al., 2011). We have submitted the paper about extra-cellular matrix(Sugiyama et al., in revision).To further investigate the roles of hepatoblasts, we used the mutant mice lacking both hepatoblasts and hepatocytes. We sorted out CD45(+) c-Kit(+) Sca-1(+) hematopoietic stem cells from E19. 5 fetal liver of both mutants and wild type control and transplanted into irradiated recipient mice. Three to six months after transplantation, hematopoietic reconstitution was analyzed by flowcytometry. The chimerism of wild type control was achieved to over 5%, whereas that of mutants was less than 0. 5%. Using these samples, we performed micro-array analysis and identified several candidate genes that likely regulate differentiation of hematopoietic stem cells.

1. KS-13信号分析通过KS-13数据库，我们鉴定了在红系分化过程中表达下调的Hmgn 2基因。Hmgn 2基因在白血病细胞系中的过表达导致体外红系分化的抑制（Kulkeaw等人，2012年）。造血干细胞分化调控新分子的鉴定。胎肝由造血细胞、肝母细胞和血窦内皮细胞组成。通过流式细胞术，我们分选出Dlk-1（+）成肝细胞，其除了造血细胞因子SCF和EPO之外，还主要表达细胞外基质如纤连蛋白和玻连蛋白（Sugiyama et al.，2011年）。我们已经提交了关于细胞外基质的论文（Sugiyama等人，为了进一步研究成肝细胞的作用，我们使用了缺乏成肝细胞和肝细胞的突变小鼠。我们从E19中分选出CD 45（+）c-Kit（+）Sca-1（+）造血干细胞。将突变体和野生型对照的5个胎肝移植到辐射受体小鼠中。移植后3 ~ 6个月，用流式细胞术分析造血重建情况。野生型对照的嵌合率达到5%以上，而突变体的嵌合率低于0。百分之五使用这些样本，我们进行了微阵列分析，并确定了几个候选基因，可能调控造血干细胞的分化。