Cryopreservation of human kidney organoids

• DOI: 10.1007/s00018-024-05352-7
• Publication Date: 2024-07-18T11:01:37Z
• Authors: Parham Mashouf; Nahid Tabibzadeh; Shohei Kuraoka; Haruka Oishi; Ryuji Morizane
• Keywords: Organoid; Vitrification; Cryoprotectant

Recent advances in stem cell research have led to the creation of organoids, miniature replicas human organs, offering innovative avenues for studying diseases. Kidney with their ability replicate complex renal structures, provide a novel platform investigating kidney diseases and assessing drug efficacy, albeit hindered by labor-intensive generation batch variations, highlighting need tailored cryopreservation methods enable widespread utilization. Here, we evaluated strategies organoids contrasting slow-freezing vitrification methods. 118 were categorized into five conditions. Control followed standard culture, while two groups used 10% DMSO (SF1) or commercial freezing media (SF2). Vitrification involved V1 (20% DMSO, 20% Ethylene Glycol sucrose) V2 (15% 15% Glycol). Assessment viability, functionality, structural integrity post-thawing revealed notable differences. Vitrification, particularly V1, exhibited superior viability (91% 26% V2, 79% SF1, 83% SF2 compared 99.4% controls). 3D imaging highlighted distinct nephron segments among groups, emphasizing V1's efficacy preserving both podocytes tubules organoids. Cisplatin-induced injury significant reduction regenerative capacities cryopreserved flow-freezing methods, method did not show statistical significance unfrozen controls. This study underscores vitrification, especially high concentrations cryoprotectants, as an effective approach maintaining organoid structure during cryopreservation, practical approaches research.