人脑器官样本为人类脑发育和病理模拟提供了独特平台。然而，目前的脑器官系统大多数缺乏分辨率以重现具有亚区域识别能力的更精细的脑结构发育，包括丘脑内功能不同的核。在这里，我们报道了一种将人类胚胎干细胞(hESCs)转化成带有转录多样性核身份的腹侧丘脑器官样本(vThOs)的方法。值得注意的是，单细胞RNA测序揭示了前所未有的丘脑编排，其中包括丘脑网状核（TRN）特征，这是位于腹侧丘脑的GABA能核。利用vThOs，我们探索了与TRN特异性、与疾病相关的基因ptched domain containing 1 (PTCHD1)和受体酪氨酸激酶（ERBB4）在人丘脑发育过程中的功能。PTCHD1或ERBB4的扰动虽然没有影响整个丘脑发育，但却损害了vThOs中的神经元功能。总之，vThOs提供了一个实验模型，用于理解人类脑丘脑核特异性发育和病理学。版权所有©2023 Elsevier Inc.。保留所有权利。

Human brain organoids provide unique platforms for modeling several aspects of human brain development and pathology. However, current brain organoid systems mostly lack the resolution to recapitulate the development of finer brain structures with subregional identity, including functionally distinct nuclei in the thalamus. Here, we report a method for converting human embryonic stem cells (hESCs) into ventral thalamic organoids (vThOs) with transcriptionally diverse nuclei identities. Notably, single-cell RNA sequencing revealed previously unachieved thalamic patterning with a thalamic reticular nucleus (TRN) signature, a GABAergic nucleus located in the ventral thalamus. Using vThOs, we explored the functions of TRN-specific, disease-associated genes patched domain containing 1 (PTCHD1) and receptor tyrosine-protein kinase (ERBB4) during human thalamic development. Perturbations in PTCHD1 or ERBB4 impaired neuronal functions in vThOs, albeit not affecting the overall thalamic lineage development. Together, vThOs present an experimental model for understanding nuclei-specific development and pathology in the thalamus of the human brain.Copyright © 2023 Elsevier Inc. All rights reserved.