Directed differentiation of mouse embryonic stem cells into thyroid follicular cells
Directed differentiation of mouse embryonic stem cells into thyroid follicular cells
复制标题
DOI：
10.1210/en.2005-1239
复制
发表时间：
2006-06-01
期刊：
影响因子：
4.8
通讯作者：
Lin, Reigh-Yi
中科院分区：
文献类型：
--
作者：
Arufe, Maria C.;Lu, Min;Lin, Reigh-Yi
阐明导致甲状腺滤泡细胞诱导和分化的分子机制对于我们了解甲状腺发育非常重要。为了表征这一过程中的关键事件，我们之前建立了实验性胚胎干（ES）细胞模型系统，该系统表明野生型小鼠CCE ES细胞可以在体外产生甲状腺细胞样细胞。我们通过使用基因操纵的 ES 细胞系扩展了本报告中的分析，其中绿色荧光蛋白 (GFP) cDNA 靶向 TSH 受体 (TSHR) 基因，将 GFP 表达与内源 TSHR 基因的转录联系起来。 GFP 阳性细胞的出现取决于未分化 ES 细胞形成胚状体，并且在分化的前 2-4 天通过 TSH 处理大大增强。在 Matrigel 的支持下，高度富集的 ES 细胞衍生的 GFP 阳性细胞在补充有 TSH 的无血清培养基中形成甲状腺滤泡样簇。重要的是，这些簇显示了甲状腺滤泡细胞的特征。免疫荧光研究证实了 TSHR 与 Na+/I- 同向转运蛋白在簇中的共定位，并表明 Na+/I- 同向转运蛋白仅在质膜中表达。此外，在这些细胞中观察到 I- 摄取活性。我们的结果表明ES细胞可以被诱导分化为甲状腺滤泡细胞，为研究胚胎甲状腺发育和功能提供了有力的工具。
Elucidating the molecular mechanisms leading to the induction and specification of thyroid follicular cells is important for our understanding of thyroid development. To characterize the key events in this process, we previously established an experimental embryonic stem (ES) cell model system, which shows that wild-type mouse CCE ES cells can give rise to thyrocyte-like cells in vitro. We extend our analysis in this report by using a genetically manipulated ES cell line in which green fluorescent protein (GFP) cDNA is targeted to the TSH receptor (TSHR) gene, linking GFP expression to the transcription of the endogenous TSHR gene. The appearance of GFP-positive cells was dependent on the formation of embryoid bodies from undifferentiated ES cells and was greatly enhanced by TSH treatment during the first 2-4 d of differentiation. With the support of Matrigel, highly enriched ES cell-derived GFP-positive cells formed thyroid follicle-like clusters in a serum-free medium supplemented with TSH. Importantly, these clusters display the characteristics of thyroid follicular cells. Immunofluorescent studies confirmed the colocalization of TSHRwith the Na+/I- symporter in the clusters and indicated that Na+/I- symporter was expressed exclusively in the plasma membrane. In addition, I- uptake activity was observed in these cells. Our results indicate that ES cells can be induced to differentiate into thyroid follicular cells, providing a powerful tool to study embryonic thyroid development and function.