Sofia Cristiani , Andrea Bertolini , Vittoria Carnicelli , Lucia Contu , Valentina Vitelli , Alessandro Saba , Federica Saponaro , Grazia Chiellini , Antonietta Raffaella Maria Sabbatini , Maria Anita Giambelluca , Paola Lenzi , Francesco Fornai , Leonardo Rossi , Gabriele Materazzi , Carlo Enrico Ambrosini , Grazia Rutigliano , Riccardo Zucchi , Ranieri Bizzarri , Sandra Ghelardoni
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引用次数: 0
摘要
为了解决二维培养的局限性,并研究 3-MNT 等化合物对脱卤酶 1(IYD)和代谢活动的影响,我们开发了一种三维甲状腺模型。在有或没有促甲状腺激素刺激和 3-MNT 处理的情况下,用 TEM 评估了形态学,并用 qRT-PCR、免疫荧光、Western 印迹、ELISA 和 LC-MS/MS 分析了组织特异性基因(TPO、TSHR、PAX8、TTF-1、NIS、IYD、TG)的表达和代谢特征。共聚焦和 TEM 分析证实了滤泡样三维结构。与二维培养物相比,三维培养物中 TPO、NIS、TG、TSH 和 PAX 标记物的表达量明显更高(p < 0.05),ELISA 显示 TG 蛋白生成量增加。3-MNT 处理可抑制 IYD 活性(表现为培养基中 MIT 和 DIT 的增加),并显著改变 NIS、TG、IYD、TSHR 和 TPO 的表达(p < 0.05)。这些研究结果表明,三维甲状腺培养物密切复制了组织的特征和功能,为甲状腺研究提供了一种有价值的工具。
Development and primary characterization of a human thyroid organoid in vitro model for thyroid metabolism investigation
A 3D thyroid model was developed to address the limitations of 2D cultures and study the effects of compounds like 3-MNT on dehalogenase 1 (IYD) and metabolic activity. Morphology was assessed by TEM, and the expression of tissue-specific genes (TPO, TSHR, PAX8, TTF-1, NIS, IYD, TG) and metabolic features were analyzed using qRT-PCR, immunofluorescence, western blotting, ELISA, and LC-MS/MS, with and without TSH stimulus and 3-MNT treatment. Confocal and TEM analyses confirmed a follicle-like 3D structure. Expression of TPO, NIS, TG, TSH, and PAX markers was significantly higher (p < 0.05) in 3D versus 2D cultures, and ELISA showed increased TG protein production. 3-MNT treatment inhibited IYD activity, indicated by increased MIT and DIT in the media, and significantly altered (p < 0.05) NIS, TG, IYD, TSHR, and TPO expression. These findings suggest 3D thyroid cultures closely replicate tissue traits and functionality, providing a valuable tool for thyroid research.
期刊介绍:
Molecular and Cellular Endocrinology was established in 1974 to meet the demand for integrated publication on all aspects related to the genetic and biochemical effects, synthesis and secretions of extracellular signals (hormones, neurotransmitters, etc.) and to the understanding of cellular regulatory mechanisms involved in hormonal control.