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原文来源自“网络期刊PLoS Current---以开放性获取期刊闻名的PLoS学术组织”
3 s8 q2 M% y8 }7 u; P由干细胞之家新闻小组成员Hyde翻译（转帖请注明）6 {8 t% K c4 f/ G
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在不形成胚体的情况下将鼠ESC分化形成内胚层& A. R( D8 m# j. D h( C! b2 i" ]
摘要：
; V9 s* @# Q! D6 R, m/ y8 I% C具有多能性的ESC在治疗一些慢性组织器官疾病（如1型糖尿病等）是一个潜在而又庞大的替代性组织器官的来源。因此，我们设计一个实验操作过程，使用全反式维甲酸、碱性成纤维细胞生长因子、丁酰环磷酸腺苷，在没有胚体形成的情况下，将鼠的ESC分化为内胚层组织，该组织可以作为胰腺前提。所生产的细胞进行定量PCR分析、免疫组织化学、静态胰岛素释放实验监测胚胎三胚层细胞、胰脏细胞的特异性标记。分化后细胞的Sox17、Foxa2表达增加，符合内胚层细胞的特点。Sox7即一种胚外内胚层细胞的标志物和Oct4即一种多能性细胞的标志物，它们的量很少。中胚层和神经外胚层形成时分别会表达的一定水平的brachyury 和Sox1的量也很少。各项检查指标显示这一实验过程所产生的细胞群体表达胰系细胞的标记性物质，其中包括胰岛素I、胰岛素II、c-肽、PDX-1、羧肽酶E、泛细胞角蛋白、淀粉酶、胰高血糖素、PAX6、Ngn3、Nkx6.1。在实验过程中我们使用了全反式维甲酸、丁酰环磷酸腺苷，没有形成胚体，生成的细胞群具有明明显的内胚层细胞特点，可作为胰腺前体候选来源。: u9 s8 x" n5 ]9 M8 _# z! [) f1 v
Differentiation of Mouse Embryonic Stem Cells into Endoderm without Embryoid Body Formation
; @) x; J7 ^( `Abstract
0 W8 q( F1 p3 Q+ s$ L- _4 M0 YPluripotent embryonic stem cells hold a great promise as an unlimited source of tissue for treatment of chronic diseases such as Type 1 diabetes. Herein, we describe a protocol using all-trans-retinoic acid, basic fibroblast growth factor and dibutyryl cAMP (DBcAMP) in the absence of embryoid body formation, for differentiation of murine embryonic stem cells into definitive endoderm that may serve as pancreatic precursors. The produced cells were analyzed by quantitative PCR, immunohistochemistry and static insulin release assay for markers of trilaminar embryo, and pancreas. Differentiated cells displayed increased Sox17 and Foxa2 expression consistent with definitive endoderm production. There was minimal production of Sox7, an extraembryonic endoderm marker, and Oct4, a marker of pluripotency. There was minimal mesoderm or neuroectoderm formation based on expression levels of the markers brachyury and Sox1, respectively. Various assays revealed that the cell clusters generated by this protocol express markers of the pancreatic lineage including insulin I, insulin II, C-peptide, PDX-1, carboxypeptidase E, pan-cytokeratin, amylase, glucagon, PAX6, Ngn3 and Nkx6.1. This protocol using all-trans-retinoic acid, DBcAMP, in the absence of embryoid bodies, generated cells that have features of definitive endoderm that may serve as pancreatic endocrine precursors.9 n' m1 Z2 C) a' a; b
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PS：个人翻译，理解不准确的语句还望大家积极指出。有些专业名称翻译不当请见谅。
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http://www.plosone.org/article/i ... ournal.pone.0014146;jsessionid=31F352661B487E628167CA2B1EE802B0.ambra02; L. Q# U6 x1 u. n1 b6 v
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发表于 2010-12-5 12:04
