Work

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

Year: 2025

Type: article

Abstract: 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

Source: STAR Protocols

DOI: 10.1016/j.xpro.2025.103983

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

Institutions: Zhongda Hospital Southeast University, State Key Laboratory of Digital Medical Engineering, Southeast University, Nanjing Medical University

Language: English

Topic: Pluripotent Stem Cells Research

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: gold

Estimated APC paid: $1,550