将人多能干细胞分化为中脑类器官用于病毒靶向显微注射的方案。

IF 1.3 Q4 BIOCHEMICAL RESEARCH METHODS

STAR Protocols Pub Date : 2025-07-25 DOI:10.1016/j.xpro.2025.103983

Mengdan Tao, Qi Chen, WeiWei Gao, Shanshan Wu, Yan Liu

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引用次数: 0

摘要

中脑类器官是研究大脑发育和疾病的有价值的模型。在这里，我们提出了一种中脑类器官模型的病毒感染方案。我们描述了人类多能干细胞（hPSC）向中脑类器官分化的步骤；设备准备;并将病毒靶向注射到类器官中，以减少病毒载量，最大限度地减少细胞毒性，并保持结构完整性。然后详细说明观察感染细胞的程序。通过将病毒与特定启动子结合，该方案能够精确靶向管状心室区（VZ）区域。有关本协议使用和执行的完整细节，请参见Zhu等人，1 Wu等人，2 Xu等人，3和Lu等人。

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Protocol for differentiating human pluripotent stem cells into midbrain organoids for targeted microinjection of viruses.

Midbrain organoids serve as a valuable model for studying brain development and disease. Here, we present a viral infection protocol for midbrain organoid models. We describe steps for human pluripotent stem cell (hPSC) differentiation into midbrain organoids; equipment preparation; and targeted virus microinjection into organoids to reduce viral load, minimize cytotoxicity, and preserve structural integrity. We then detail procedures for visualizing infected cells. By combining the virus with specific promoters, this protocol enables precise targeting of tube-like ventricular zone (VZ) regions. For complete details on the use and execution of this protocol, please refer to Zhu et al.,¹ Wu et al.,² Xu et al.,³ and Lu et al.⁴.

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