Effective cryopreservation of human brain tissue and neural organoids

• Published in: Cell reports methods Vol. 4; no. 5; p. 100777
• Main Authors: Xue, Weiwei, Li, Huijuan, Xu, Jinhong, Yu, Xiao, Liu, Linlin, Liu, Huihui, Zhao, Rui, Shao, Zhicheng
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 20.05.2024; Elsevier
• Subjects: brain; brain diseases; cryopreservation; ethylene glycol; living brain tissue; methylcellulose; neural organoid; methylcellulose; living brain tissue; brain diseases; ethylene glycol; neural organoid; CP: Stem cell; brain; cryopreservation
• ISSN: 2667-2375; 2667-2375
• DOI: 10.1016/j.crmeth.2024.100777

Human brain tissue models and organoids are vital for studying and modeling human neurological disease. However, the high cost of long-term cultured organoids inhibits their wide-ranging application. It is therefore urgent to develop methods for the cryopreservation of brain tissue and organoids. Here, we establish a method using methylcellulose, ethylene glycol, DMSO, and Y27632 (termed MEDY) for the cryopreservation of cortical organoids without disrupting the neural cytoarchitecture or functional activity. MEDY can be applied to multiple brain-region-specific organoids, including the dorsal/ventral forebrain, spinal cord, optic vesicle brain, and epilepsy patient-derived brain organoids. Additionally, MEDY enables the cryopreservation of human brain tissue samples, and pathological features are retained after thawing. Transcriptomic analysis shows that MEDY can protect synaptic function and inhibit the endoplasmic reticulum-mediated apoptosis pathway. MEDY will enable the large-scale and reliable storage of diverse neural organoids and living brain tissue and will facilitate wide-ranging research, medical applications, and drug screening. [Display omitted] •We establish a cryopreservation method (MEDY) for brain organoids and tissues•MEDY maintains the structure and functional activity of cortical organoids•MEDY can be applied to diverse brain organoids, including patient-derived organoids Brain organoids are useful models for modeling brain diseases. However, long-term culture, high costs, and storage problems significantly limit their application in biomedical studies. Here, we establish a method using methylcellulose, ethylene glycol, DMSO, and Y27632 (termed MEDY) for the cryopreservation of diverse brain organoids or human brain tissue without disrupting the neural cytoarchitecture or functional activity. Xue et al. establish a brain organoid cryopreservation technique termed MEDY. MEDY allows cryopreservation without disrupting the neural cytoarchitecture and is applicable to diverse brain organoids, including those derived from patients or long-term culture, as well as damaged brain tissue.