Development of efficient hematopoietic stem cell proliferation systems using our own established ES cells of small monkey, common marmoset

使用我们自己建立的小猴、狨猴的ES细胞开发高效的造血干细胞增殖系统

基本信息

批准号：
17390279
负责人：
TANI Kenzaburo
金额：
$ 9.86万
依托单位：
KYUSHU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2006
项目状态：
已结题

项目摘要

Toward the goal of establishing efficient hematopoietic stem cells and hematopoietic differentiation system from our own established common marmoset(CM) embryonic stem(ES) cells, we first established tall/scl gene transduced CMES (tal1/scl) cell lines. Tall/scl gene transduction was found to differentiate CMES cells efficiently to hematopoietic cells in vitro in our previous studies. We first of all found in vitro differentiating capacity of the tal1/scl cells to lymphocytes and megakaryocytes, although the efficiency was low. On the other hand, our in vivo studies using immune deficient NOG mice demonstrated very poor hematopoietic reconstitution with intramarrow injected tall/scl cells because of early lung tumor production and death. Then we tried to find out second gene which may assist the hematopoietic differentiation of tall/scl cells in vivo. For this purpose, we constructed human fetal liver cDNA expression VSV-G pseudotyped lentiviral vector library. This library consisted of more than 8x10^7 individual clones with average cDNA length of 2.1kb. DNA sequencing analysis of several fractions showed 60% of them contained full length cDNAs. When we produced virus using the library with 293T cells, 100% of cells demonstrated gene transduction. Average length of the gene transduced cDNA was about 1 kb. Flow cytometric analysis with antiglycophorin antibody showed the gene transduced cells expressed glycophorin, which is known to be one of the highly abundantly expressed genes in human fetal liver, properly. Using this novel human fetal liver cDNA library, we have already cloned several candidate genes which promotes hematopoiesis of tall/scl cells or induces cytokine independency of cytokine dependent leukemia cell lines. By the identification of genes which differentiate ES cells efficiently to hematopoietic cells in vitro and in vivo, safer and more effective cell supplementation therapy would be realized in future.

目的是建立有效的造血干细胞和造血分化系统，从我们自己已建立的普通果果（CM）胚胎干细胞（ES）细胞中，我们首先建立了高/SCL基因转导的CMES（TAL1/SCL）细胞系。在我们先前的研究中，发现高/SCL基因转导可以在体外有效地将CMES细胞有效地分化为造血细胞。我们首先发现TAL1/SCL细胞在体外分化能力对淋巴细胞和巨核细胞的能力，尽管效率较低。另一方面，我们使用免疫缺陷NOG小鼠的体内研究表明，由于早期的肺肿瘤产生和死亡，造血的造血重建非常不良。然后，我们试图找出第二个基因，该基因可能有助于体内高高/SCL细胞的造血分化。为此，我们构建了人类胎儿肝cDNA表达VSV-G拟型慢病毒载体文库。该库由超过8x10^7的单个克隆组成，平均cDNA长度为2.1kb。对几个部分的DNA测序分析显示，其中60％包含全长cDNA。当我们使用带有293T细胞的文库产生病毒时，100％的细胞表现出基因转导。基因转导的cDNA的平均长度约为1 kb。用抗糖蛋白抗体的流式细胞仪分析表明，基因转导的细胞表达了糖蛋白，已知是人类胎儿肝脏中高度表达的基因之一。使用这个新型的人类胎儿肝cDNA文库，我们已经克隆了几个候选基因，该基因促进了高/SCL细胞的造血或诱导细胞因子依赖性白血病细胞系的细胞因子独立性。通过鉴定在体外和体内将ES细胞有效地与造血细胞有效分化的基因，将来将来会实现更安全，更有效的细胞补充疗法。

项目成果

期刊论文数量（0）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Serial analysis of gene expression in progressing and regressing mouse tumors implicates the involvement of RANTES and TARC in antitumor immune responses.

对进展和消退小鼠肿瘤中基因表达的系列分析表明 RANTES 和 TARC 参与抗肿瘤免疫反应。

DOI：
发表时间：
2006
期刊：
Mol Ther 14
影响因子：
0
作者：
Nakazaki;Y.;Tani;K.;et al.
通讯作者：
et al.

Sustained molecular remission by non-myeloablative stem cell transplantation after autologous hematopoietic stem cell transplantation in a patient with multiple myeloma.

多发性骨髓瘤患者在自体造血干细胞移植后通过非清髓性干细胞移植实现持续分子缓解。

DOI：
发表时间：
2005
期刊：
Leuk Lynphoma 46
影响因子：
0
作者：
Nakashima;Y.;Tani;K.;et al.
通讯作者：
et al.

Nonhuman Primates : Embryonic stem cells and transgenesis.

非人类灵长类动物：胚胎干细胞和转基因。

DOI：
发表时间：
2006
期刊：
Cloning and Stem Cells 8
影响因子：
0
作者：
Horn;P.A.;Tani;K.;et al.
通讯作者：
et al.

Rb plays a role in survival of Abl-dependent human tumor cells as a downstream effector of Abl tyrosine kinase

DOI：
10.1038/sj.onc.1208996
发表时间：
2006-01
期刊：
Oncogene
影响因子：
8
作者：
K. Nagano;C. Itagaki;T. Izumi;Kazuto Nunomura;Yasushi Soda;Kenzaburo Tani;Nobuhiro Takahashi;Tadaomi Takenawa;Toshiaki Isobe
通讯作者：
K. Nagano;C. Itagaki;T. Izumi;Kazuto Nunomura;Yasushi Soda;Kenzaburo Tani;Nobuhiro Takahashi;Tadaomi Takenawa;Toshiaki Isobe

Establishment of novel embryonic stem cell lines derived from the common marmoset (Callithrix jacchus)