Protocol for cryopreserving neural organoids and human living brain tissue using MEDY
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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
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.
Abstract:
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.
Insights
We developed a novel cryopreservation protocol using methylcellulose, ethylene glycol, DMSO, and Y27632 (MEDY) for reliable storage of neural organoids and human brain tissue, aiding neurological disease research.
Area of Science:
- Neuroscience
- Cryobiology
- Biotechnology
Background:
- Cryopreservation of neural organoids and human brain tissue is crucial for research but faces significant challenges.
- Existing methods often result in compromised tissue viability and functionality.
Purpose of the Study:
- To develop and present a reliable cryopreservation protocol for diverse neural organoids and human living brain tissue.
- To establish a standardized method for long-term storage and subsequent use of neural tissues for research.
Main Methods:
- The protocol, termed MEDY, utilizes a specific combination of methylcellulose, ethylene glycol, DMSO, and Y27632.
- Detailed procedures cover solution preparation, organoid/tissue preparation, cryopreservation, resuscitation, and detection assays.
Main Results:
- The MEDY protocol enables reliable storage of various neural organoids and human brain tissue.
- Successful cryopreservation and resuscitation were demonstrated, maintaining tissue integrity for downstream applications.
Conclusions:
- The MEDY protocol offers a robust solution for cryopreserving neural organoids and human brain tissue.
- This advancement provides a valuable model for studying human neurological diseases and facilitates broader research accessibility.
