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1. 发明申请

US20100184212A1 HUMAN EMBRYONIC STEM CELL DERIVED MESODERM-LIKE EPITHELIUM TRANSITIONS TO MESENCHYMAL PROGENITOR CELLS 审中-公开
标题翻译：人胚胎干细胞衍生的MESODERM型上皮细胞转染到细胞祖细胞
公开(公告)号：US20100184212A1
公开(公告)日：2010-07-22
申请号：US12451720
申请日：2008-05-30
申请人： Steven Stice , Nolan Boyd
发明人： Steven Stice , Nolan Boyd
IPC分类号： C12N5/0735 , C12N5/0789 , C12N5/077
CPC分类号： C12N5/0662 , C12N5/0625 , C12N2501/105 , C12N2501/11 , C12N2501/115 , C12N2501/165 , C12N2506/02
摘要： Human embryonic stem cells (hESC) have the potential to produce all of the cells in the body. They are also able to self-renew indefinitely, sparking the hope they could be used as a source for large scale production of therapeutic cell lines. The present invention relates to a monolayer differentiation culture system that induces hESC (WA09 and BG01) to form epithelial sheets with mesodermal gene expression patterns (BMP4, RUNX1, GAT A4). These E-cadherin+ CD90lovv cells then undergo apparent epithelial-mesenchymal transformation (EMT) for the derivation of mesenchymal progenitor cells (hES-MC) that by flow cytometry are negative for hematopoietic (CD34, CD45 and CD 133) and endothelial (CD31 and CD 146) markers, but positive for markers associated with mesenchymal stem cells (MSC) (CD73, CD90, CD105 and CD166). To determine their functionality, we tested their capacity to produce the three lineages commonly associated with MSC and found they could form osteogenic and chondrogenic, but not adipogenic lineages. The derived hES-MC were able to remodel and contract collagen I lattice constructs to an equivalent degree as keloid fibroblast control cells and were induced to express αSMA when exposed to TGF-β1, but not PDGF-B. This data suggests the derived hES-MC cells are multipotent cells with potential uses in tissue engineering/regenerative medicine and for providing a highly reproducible cell source for adult-like progenitor cells.
摘要翻译：人类胚胎干细胞（hESC）具有产生体内所有细胞的潜力。他们也能够无限期地自我更新，激发了他们希望将其用作大规模生产治疗性细胞系的来源。本发明涉及诱导hESC（WA09和BG01）形成具有中胚层基因表达模式（BMP4，RUNX1，GAT A4）的上皮片的单层分化培养系统。这些E-钙粘蛋白+ CD90lovv细胞然后经历明显的上皮 - 间质转化（EMT），用于衍生间充质祖细胞（hES-MC），其通过流式细胞术对造血（CD34，CD45和CD133）和内皮（CD31和CD 146）标记，但与间充质干细胞（MSC）（CD73，CD90，CD105和CD166）相关标志物阳性。为了确定其功能，我们测试了其产生通常与MSC相关的三个谱系的能力，发现它们可以形成成骨和软骨形成，但不能形成脂肪形成的谱系。衍生的hES-MC能够将胶原I晶格构建体重塑和收缩至瘢痕疙瘩成纤维细胞对照细胞的相当程度，并且当暴露于TGF-β1而不是PDGF-B时被诱导表达αSMA。该数据表明衍生的hES-MC细胞是在组织工程/再生医学中具有潜在用途的多能细胞，并为成年样祖细胞提供高度可重复的细胞来源。

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