权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Reprogramming and somatic memory in ES cell/hematopoietic stem cell hybrids

ES细胞/造血干细胞杂交体中的重编程和体细胞记忆

基本信息

批准号：
66066327
负责人：
Professor Dr. Martin Zenke
金额：
--
依托单位：
Klinik für Onkologie, Hämatologie und Stammzelltransplantation (Med. Klinik IV)
依托单位国家：
德国
项目类别：
Priority Programmes
财政年份：
2008
资助国家：
德国
起止时间：
2007-12-31 至 2015-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/66066327?language=en
关键词：
Reprogramming somatic memory ES cell

项目摘要

In this project we study somatic memory in pluripotent cells derived from hematopoietic stem cells (HSC) by (i) fusion with embryonic stem cells (ESC) and (ii) transduction with reprogramming transcription factors. Flt3+ HSC from bone marrow of Oct4-eGFP transgenic mice were fused with ESC to generate Oct4-eGFP+ reprogrammed Flt3 HSC/ESC hybrids. These Flt3 ESC hybrids were pluripotent, showed ESC morphology and an ESC-like gene expression profile, but surprisingly contained somatic memory. Flt3+ HSC from Oct4-eGFP transgenic mice were transduced with the four reprogramming factors Oct4, Sox2, c-Myc and Klf4 to obtain Oct4- eGFP+ induced pluripotent stem cells (Flt3 iPS cells). Pluripotency and somatic memory in Flt3 ESC hybrids and Flt3 iPS cells were studied in in vitro culture systems and following transfer into recipient mice in vivo. We found that Flt3 ESC hybrids with somatic memory differentiated in embryoid body (EB) assays with accelerated kinetics and reverted from 4n to 2n chromosome stage. Additionally, such EB exhibited a well-structured tissue organisation similar to a developing embryo. Flt3 iPS cells showed delayed differentiation kinetics and a moderate differentiation potential. Our studies focus now on obtaining detailed insights into the molecular basis of somatic memory in Flt3 ESC hybrids and the 4n to 2n transition observed during their differentiation.

在这个项目中，我们通过(i)与胚胎干细胞（ESC）的融合和（ii）与重编程转录因子的转导，研究来自造血干细胞（HSC）的多能细胞的体细胞记忆。将来自Oct4-eGFP转基因小鼠骨髓的Flt3+ HSC与ESC融合，生成Oct4-eGFP+重编程Flt3 HSC/ESC杂种。这些Flt3 ESC杂交种是多能的，具有ESC形态和ESC样基因表达谱，但令人惊讶的是包含体细胞记忆。用四种重编程因子Oct4、Sox2、c-Myc和Klf4转导Oct4-eGFP转基因小鼠的Flt3+ HSC，获得Oct4-eGFP +诱导的多能干细胞（Flt3 iPS细胞）。研究了Flt3 ESC杂交种和Flt3 iPS细胞的体外培养和体内移植的多能性和体细胞记忆。我们发现具有体细胞记忆的Flt3 ESC杂种在胚状体（EB）实验中以加速动力学的方式分化，并从4n染色体阶段恢复到2n染色体阶段。此外，这种EB表现出类似于发育中的胚胎的结构良好的组织组织。Flt3 iPS细胞表现出延迟分化动力学和中等分化潜力。我们现在的研究重点是获得Flt3 ESC杂交种体细胞记忆的分子基础，以及在分化过程中观察到的4n到2n的转变。