Establishment of differentiation protocols for mouse embryonic stem cells into hepatocytes and insulin producing islet β cells.

建立小鼠胚胎干细胞分化为肝细胞和产生胰岛素的胰岛β细胞的方案。

基本信息

  • 批准号:
    18591410
  • 负责人:
  • 金额:
    $ 2.51万
  • 依托单位:
  • 依托单位国家:
    日本
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
  • 财政年份:
    2006
  • 资助国家:
    日本
  • 起止时间:
    2006 至 2007
  • 项目状态:
    已结题

项目摘要

(1) Establishment of regulatory system for the expression of transcription factors in mouse ES cellsWe produced regulatory plasmid designed to express transcription factors specific for mesoendodermal cells such as Mix and Hex, which were subcloned from embryoid bodies. These transcription factors were expressed under the control of Tet-ON system. The target cells in which transcription factors were preferentially expressed could be determined as green fluorescent protein (GFP) expressing cells because GFP coding sequence was ligated after the sequence of internal ribosomal entry site (IRES). Transgenic mouse ES cells were produced after the transfection of this plasmid.(2) In vitro differentiation of mouse ES cells into mature hepatocytes and insulin producing islet β cellsAfter the isolation of GFP positive cells derived from transgenic mouse ES cells produced as previously described, which express Mix by the administration of Doxcycline in the culture medium, GFP positive cells were … More cultured under several conditions identified by the combination of several growth factors (HGF, FGF2, ATRA) and also several extra-cellular matrices (Collagen type I, IV, Laminin, Matrigel). RT-PCR was performed to identify the expression of mesodermal and endodermal marker genes in the cells during the differentiation culture. Although the expression of several mesodermal markers such as Gooscoid and brachury and also several endodermal marker such as GATA4 and FoxA2 were identified in the isolated cells, the expression of hepatocyte marker such as alfa-fetoprotein, albumin were not identified. In addition, the expression of Glut-2 and Pdx-1 were not determined whereas the expression of insulin was detected in the isolated Mix expressing cells. Therefore, the sufficient differentiation findings for hepatocytes and also insulin producing islet β cells were not comfirmed in our experiments. On the other hand, we identified the expression of hepatocyte markers in the cells derived from mouse ES cells co-cultured with Thy-1 positive mesenchymal cells originated from mouse fetal liver. We also identified morphological features as hepatocytes in these differentiated cells using phase contrast microscopy and also electron microscopy.(3) The transplantation of hepatocyte like cells derived from mouse ES cellsWe performed transplantation experiments of hepatocyte-like cells differentiated from mouse ES cells marked with LacZ gene into transgenic mice in which lethal liver damage occurred by the administration of diphtheria toxin. We identified not only the incorporation of transplanted hepatocyte like cells into recipient liver but also the improvement of mortality rate ofrecipient mice suffered from lethal liver damage. Less
(1)小鼠胚胎干细胞转录因子表达调控体系的建立我们从胚胎样体亚克隆制备了用于表达Mix和Hex等中胚层细胞特异性转录因子的调控质粒。这些转录因子在Tet-ON系统的调控下表达。转录因子优先表达的靶细胞可以确定为绿色荧光蛋白(GFP)表达细胞,因为GFP编码序列连接在内部核糖体进入位点(IRES)序列之后。转染该质粒后,获得了转基因小鼠胚胎干细胞。(2)小鼠胚胎干细胞体外分化为成熟肝细胞和产生胰岛素的胰岛β细胞。在培养基中加入多西环素表达Mix的转基因小鼠胚胎细胞中分离出GFP阳性细胞后,在几种生长因子(HGF、FGF2、FGF2、FGF2)联合鉴定的几种条件下培养出GFP阳性细胞。ATRA)和一些细胞外基质(胶原I型,IV型,层粘连蛋白,基质)。RT-PCR检测分化培养过程中细胞中胚层和内胚层标记基因的表达情况。虽然在分离的细胞中检测到Gooscoid和brachury等中胚层标志物以及GATA4和FoxA2等内胚层标志物的表达,但未检测到肝细胞标志物如甲胎蛋白、白蛋白的表达。此外,未检测Glut-2和Pdx-1的表达,而在分离的Mix表达细胞中检测到胰岛素的表达。因此,我们的实验没有充分证实肝细胞和产生胰岛素的胰岛β细胞的分化结果。另一方面,我们在小鼠胚胎干细胞与来自小鼠胎肝的Thy-1阳性间充质细胞共培养的细胞中鉴定了肝细胞标志物的表达。我们还利用相差显微镜和电子显微镜鉴定了这些分化细胞的肝细胞形态特征。(3)小鼠胚胎干细胞衍生肝细胞样细胞的移植我们将标记有LacZ基因的小鼠胚胎干细胞分化成的肝细胞样细胞移植到白喉毒素介导的转基因小鼠中,对小鼠的肝造成致死性损伤。我们不仅发现了移植的肝细胞样细胞与受体肝脏的结合,而且还发现了遭受致死性肝损伤的受体小鼠死亡率的改善。少

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)
Transplantation of embryonic stem cell-derived endodermal cells into mice with life-threatning liver
将胚胎干细胞衍生的内胚层细胞移植到肝脏危及生命的小鼠体内
  • DOI:
  • 发表时间:
    2007
  • 期刊:
  • 影响因子:
    0
  • 作者:
    Ishii T;et. al.
  • 通讯作者:
    et. al.
Improvement of survival rate of lethally liver-injured model mice after cell transplantation of endodermal cells derived from mouse embryonic stem cells
小鼠胚胎干细胞来源的内胚层细胞移植后致死性肝损伤模型小鼠存活率的提高
  • DOI:
  • 发表时间:
    2007
  • 期刊:
  • 影响因子:
    0
  • 作者:
    lsnu l;YasuchikaK;et. al.
  • 通讯作者:
    et. al.
Improvement of survival rate of lethal liver damaged model mice after cell transplantation of endodermeal cells derived from mouse embryonic stem cells
小鼠胚胎干细胞内胚层细胞移植后致死性肝损伤模型小鼠存活率的提高
  • DOI:
  • 发表时间:
    2007
  • 期刊:
  • 影响因子:
    0
  • 作者:
    Ishii T;Yasuchika K;et. al.
  • 通讯作者:
    et. al.
Improvement of the survival rate by fetal liver cell transplantation in a mice lethal liver failure model
  • DOI:
    10.1097/01.tp.0000287967.54222.4d
  • 发表时间:
    2007-11-27
  • 期刊:
  • 影响因子:
    6.2
  • 作者:
    Machimoto, Takafumi;Yasuchika, Kentaro;Ikai, Iwao
  • 通讯作者:
    Ikai, Iwao
マウスES細胞由来内胚葉細胞を用いた細胞移植
使用小鼠 ES 细胞来源的内胚层细胞进行细胞移植
  • DOI:
  • 发表时间:
    2007
  • 期刊:
  • 影响因子:
    0
  • 作者:
    Takada;T.;T. Ishii;T. Machimoto;Y. Kimura;S. Tatsumi;Y. Miyake;河野 憲二;河野憲二;斉藤 美知子;新井布美子;松岡邦枝;古川賀絵;河野憲二;河野憲二;河野憲二;石井隆道
  • 通讯作者:
    石井隆道
{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ monograph.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ sciAawards.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ conferencePapers.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ patent.updateTime }}

YASUCHIKA Kentaro其他文献

YASUCHIKA Kentaro的其他文献

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

{{ truncateString('YASUCHIKA Kentaro', 18)}}的其他基金

Establishment of hepatocytes differentiation from human multipotent stem cells and investigation for the effectiveness of cell transplantation
人多能干细胞分化肝细胞的建立及细胞移植有效性的研究
  • 批准号:
    24591999
  • 财政年份:
    2012
  • 资助金额:
    $ 2.51万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Isolation and characterization of murine hepatic stem cells (HPCs) based on a new surface antigen.
基于新表面抗原的鼠肝干细胞 (HPC) 的分离和表征。
  • 批准号:
    20591610
  • 财政年份:
    2008
  • 资助金额:
    $ 2.51万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)

相似国自然基金

基于肝炎病毒嗜肝性对hESC-derived hepatocytes 体外分化过程中的关键因子分析
  • 批准号:
    81870432
  • 批准年份:
    2018
  • 资助金额:
    57.0 万元
  • 项目类别:
    面上项目
hESC-derived hepatocytes 中抗HBV干扰素反应模式及关键ISGs 的功能分析
  • 批准号:
    81570567
  • 批准年份:
    2015
  • 资助金额:
    57.0 万元
  • 项目类别:
    面上项目
骨髓间充质干细胞肝内移植分化为肝细胞的活体基因显像研究
  • 批准号:
    81070349
  • 批准年份:
    2010
  • 资助金额:
    30.0 万元
  • 项目类别:
    面上项目
ADF/cofilin去磷酸化与肝细胞极性重建
  • 批准号:
    30571763
  • 批准年份:
    2005
  • 资助金额:
    23.0 万元
  • 项目类别:
    面上项目

相似海外基金

Study into the effect of candidalysin exotoxin secretion and fungal ROS generation on C. albicans infection on human hepatocytes
念珠菌素外毒素分泌和真菌ROS生成对白色念珠菌感染人肝细胞影响的研究
  • 批准号:
    24K18435
  • 财政年份:
    2024
  • 资助金额:
    $ 2.51万
  • 项目类别:
    Grant-in-Aid for Early-Career Scientists
Targeted Delivery of Biodistribution-Guided Recombinant Adeno-associated Viral Vector (AAV) to Specific Hepatocytes
将生物分布引导的重组腺相关病毒载体 (AAV) 靶向递送至特定肝细胞
  • 批准号:
    24K18551
  • 财政年份:
    2024
  • 资助金额:
    $ 2.51万
  • 项目类别:
    Grant-in-Aid for Early-Career Scientists
Pluripotent stem cell derived hepatocytes for liver failure (PUSH for LIFE)
多能干细胞衍生的肝细胞治疗肝衰竭(PUSH for LIFE)
  • 批准号:
    MR/Z503940/1
  • 财政年份:
    2024
  • 资助金额:
    $ 2.51万
  • 项目类别:
    Research Grant
The unique and complementary functions of PCSK7 and PCSK9 in cardiometabolic health and disease
PCSK7 和 PCSK9 在心脏代谢健康和疾病中的独特和互补功能
  • 批准号:
    488392
  • 财政年份:
    2023
  • 资助金额:
    $ 2.51万
  • 项目类别:
    Operating Grants
Senescent hepatocytes mediate reprogramming of immune cells in acute liver failure
衰老肝细胞介导急性肝衰竭中免疫细胞的重编程
  • 批准号:
    10679938
  • 财政年份:
    2023
  • 资助金额:
    $ 2.51万
  • 项目类别:
Thyroid Hormone Signaling in Human Hepatocytes
人肝细胞中的甲状腺激素信号传导
  • 批准号:
    10874207
  • 财政年份:
    2023
  • 资助金额:
    $ 2.51万
  • 项目类别:
The role of MKP-1/MAPK in hepatocytes and macrophages in alcohol-associated liver disease pathogenesis
MKP-1/MAPK在肝细胞和巨噬细胞中在酒精相关性肝病发病机制中的作用
  • 批准号:
    10679716
  • 财政年份:
    2023
  • 资助金额:
    $ 2.51万
  • 项目类别:
Bioartificial interventions for organ senescence: Understanding and preventing senescence induced by liver toxins (SILT) in hepatocytes.
器官衰老的生物人工干预:了解和预防肝细胞中肝毒素(SILT)诱导的衰老。
  • 批准号:
    MR/Y010299/1
  • 财政年份:
    2023
  • 资助金额:
    $ 2.51万
  • 项目类别:
    Research Grant
Waking up Hepatitis B virus (HBV)'s chronic form to maximize attack by promoter-targeting therapies
唤醒乙型肝炎病毒 (HBV) 的慢性形式,通过启动子靶向疗法最大限度地发挥攻击作用
  • 批准号:
    486942
  • 财政年份:
    2023
  • 资助金额:
    $ 2.51万
  • 项目类别:
    Miscellaneous Programs
Mechanism for hepatitis E virus exit from polarized hepatocytes
戊型肝炎病毒从极化肝细胞中退出的机制
  • 批准号:
    10578386
  • 财政年份:
    2023
  • 资助金额:
    $ 2.51万
  • 项目类别:
{{ showInfoDetail.title }}

作者:{{ showInfoDetail.author }}

知道了