Miller Duncan C, Genehr Carolin, Telugu Narasimha S, Kurths Silke, Diecke Sebastian
Core Facility Stem Cells, Max-Delbrück-Centrum, Berlin, Germany.
DZHK (German Centre for Cardiovascular Research), partner site Berlin, Berlin, Germany.
Curr Protoc Stem Cell Biol. 2020 Dec;55(1):e125. doi: 10.1002/cpsc.125.
Great progress has been made with protocols for the differentiation and functional application of hPSC-cardiomyocytes (hPSC-CMs) in recent years; however, the cryopreservation and recovery of hPSC-CMs still presents challenges and few reports describe in detail the protocols and general workflow. In order to facilitate cryopreservation and recovery of hPSC-CMs for a wide range of applications, we provide detailed information and step-by-step protocols. The protocols are simple and use common reagents. They are comprised of a fast dissociation, cryopreservation using standard equipment, and gentle recovery following thawing. We discuss various features of the protocols, as well as their utilization in the context of common hPSC-CM differentiation and application workflows. Finally, we compare two proprietary and two common in-house formulations of cryopreservation media used for hPSC-CMs, and despite differences in their price and composition find broadly similar recovery rates and cellular function after thawing. © 2019 The Authors. Basic Protocol 1: Dissociation and cryopreservation of hPSC-CMs Basic Protocol 2: Thawing and recovery of cryogenically frozen hPSC-CMs.
近年来，人多能干细胞衍生心肌细胞（hPSC-CMs）的分化及功能应用方案取得了巨大进展；然而，hPSC-CMs的冷冻保存和复苏仍然存在挑战，很少有报告详细描述相关方案及一般工作流程。为了便于hPSC-CMs的冷冻保存和复苏以用于广泛的应用，我们提供了详细信息和分步方案。这些方案简单且使用常见试剂。它们包括快速解离、使用标准设备进行冷冻保存以及解冻后的温和复苏。我们讨论了这些方案的各种特点，以及它们在常见的hPSC-CM分化和应用工作流程中的应用。最后，我们比较了用于hPSC-CMs的两种专利和两种常见的内部冷冻保存培养基配方，尽管它们在价格和成分上存在差异，但解冻后的复苏率和细胞功能大致相似。© 2019作者。基本方案1：hPSC-CMs的解离和冷冻保存 基本方案2：低温冷冻的hPSC-CMs的解冻和复苏。
