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Updated: Jun 12, 2025
Generation of Standardized and Reproducible Forebrain-type Cerebral Organoids from Human Induced Pluripotent Stem Cells
Published on: January 23, 2018
Protocol for cryopreserving neural organoids and human living brain tissue using MEDY
Weiwei Xue1, Jiacheng Du1, Yunteng Li1
1Jinshan Hospital, Institute for Translational Brain Research, State Key Laboratory of Medical Neurobiology, MOE Frontiers Center for Brain Science, Institute of Pediatrics, National Children's Medical Center, Children's Hospital, Fudan University, Shanghai 200032, China.
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Cryopreservation of neural organoids and human living brain tissue faces huge challenges. Here, we present a cryopreservation protocol for the reliable storage of diverse neural organoids and human living brain tissue using methylcellulose, ethylene glycol, DMSO, and Y27632, termed MEDY. We describe steps for preparing cryopreservation solution, preparing cortical organoids, and surgical removal of human brain tissue. We then detail procedures for cryopreservation, resuscitation, and detection assays. This protocol can serve as a model for studying human neurological diseases. For complete details on the use and execution of this protocol, please refer to Xue et al.1.
