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Updated: Sep 13, 2025
Author Spotlight: Targeted Microinjection and Electroporation of Primate Cerebral Organoids for Genetic Modification
Published on: March 24, 2023
Protocol for differentiating human pluripotent stem cells into midbrain organoids for targeted microinjection of
Mengdan Tao1, Qi Chen2, WeiWei Gao2
1Institute for Stem Cell and Neural Regeneration, School of Pharmacy, Nanjing Medical University, Nanjing 211166, China; State Key Laboratory of Digital Medical Engineering, School of Biological Science and Medical Engineering, Department of Neurology, Affiliated Zhongda Hospital, Southeast University, Nanjing 210096, China.
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.
