Development of heart organoid cryopreservation method through Fe₃O₄ nanoparticles based nanowarming system

Abstract

Beyond single cell two‐dimensional (2D) culture, research on organoids that can mimic human organs is rapidly developing. However, there are still problems in commercialization and joint research using organoids due to the lack of technology to safely store organoids. Since organoids are 3D complex structures with a certain size (0.1 to 5 mm) beyond the size of cells, the conventional cell‐level cryopreservation method using cryoprotectant (CPA) cannot overcome the damage caused by volume change due to osmotic pressure difference and ice nucleation. Herein, we attempted to solve such limitations by applying a nanowarming system using CPA with high cell permeability and Fe3O4 nanoparticles. By performing beat rate measurement, histological analysis, contractility analysis, and multi‐electrode array, it was verified that the developed method could significantly improve functional recovery and survival of heart organoids after freezing and thawing. In this study, we demonstrated a successful organoid cryopreservation method based on a Fe3O4 nanowarming system. The developed technology will provide clues to the field of tissue cryopreservation and spur the application of organoids.This article is protected by copyright. All rights reserved

Cryopreservation of neuroectoderm on a pillar plate andin situdifferentiation into human brain organoids

“…The establishment of cryopreservation and thawing protocols for organoids could enhance biobanking and the utility of organoids in research and therapy. While several studies have investigated cryopreservation methods for 3D cell culture models, no specific protocol has yet been developed for brain organoids [21][22][23][24][25] . The cryopreservation of organoids is influenced by various factors, including mechanical stress, cooling and thawing rates, organoid size, and the type of cryoprotective agents (CPAs).…”

Cryopreservation of organoids: Strategies, innovation, and future prospects

“…Magnetic nano-rewarming has been used for vitrification preservation of cardiac organs. Lee et al [199] demonstrated a successful cryopreservation method for organoids based on a Fe 3 O 4 magnetic nano-rewarming system. It has been verified through heartrate measurement, histological analysis, contraction force analysis, and multielectrode array that magnetic nano-rewarming can significantly improve the functional recovery and survival rate of heart organs after freeze-thaw.…”