低温保护剂
球体
低温保存
二甲基亚砜
体内
细胞外
低温生物学
肝细胞
材料科学
生物物理学
细胞生物学
高分子
玻璃化
肌动蛋白
体外
化学
生物
生物化学
男科
胚胎
有机化学
生物技术
医学
作者
Akalabya Bissoyi,Ruben M F Tomás,Yanan Gao,Qiongyu Guo,Matthew I. Gibson
标识
DOI:10.1021/acsami.2c18288
摘要
Spheroids are a powerful tool for basic research and to reduce or replace in vivo (animal) studies but are not routinely banked nor shared. Here, we report the successful cryopreservation of hepatocyte spheroids using macromolecular (polyampholyte) cryoprotectants supplemented into dimethyl sulfoxide (DMSO) solutions. We demonstrate that a polyampholyte significantly increases post-thaw recovery, minimizes membrane damage related to cryo-injury, and remains in the extracellular space making it simple to remove post-thaw. In a model toxicology challenge, the thawed spheroids matched the performance of fresh spheroids. F-actin staining showed that DMSO-only cryopreserved samples had reduced actin polymerization, which the polyampholyte rescued, potentially linked to intracellular ice formation. This work may facilitate access to off-the-shelf and ready-to-use frozen spheroids, without the need for in-house culturing. Readily accessible 3-D cell models may also reduce the number of in vivo experiments.
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