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{"title":"Simple Workflow and Comparison of Media for hPSC-Cardiomyocyte Cryopreservation and Recovery","authors":"Duncan C. Miller, Carolin Genehr, Narasimha S. Telugu, Silke Kurths, Sebastian Diecke","doi":"10.1002/cpsc.125","DOIUrl":null,"url":null,"abstract":"<p>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.</p><p><b>Basic Protocol 1</b>: Dissociation and cryopreservation of hPSC-CMs</p><p><b>Basic Protocol 2</b>: Thawing and recovery of cryogenically frozen hPSC-CMs</p>","PeriodicalId":53703,"journal":{"name":"Current Protocols in Stem Cell Biology","volume":"55 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2020-09-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpsc.125","citationCount":"4","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Stem Cell Biology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpsc.125","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
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Abstract
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
hpscs -心肌细胞低温保存与恢复的简单工作流程及培养基比较
近年来，人类造血干细胞-心肌细胞(hpscs - cms)的分化和功能应用研究取得了很大进展;然而，hscs - cms的冷冻保存和恢复仍然存在挑战，很少有报道详细描述了协议和一般工作流程。为了促进超低温保存和恢复hscs - cms的广泛应用，我们提供了详细的信息和一步一步的协议。该方法简单，使用常用试剂。它们由快速分离，使用标准设备冷冻保存和解冻后的温和恢复组成。我们讨论了协议的各种特性，以及它们在通用hPSC-CM区分和应用程序工作流环境中的使用。最后，我们比较了两种专有和两种常见的用于hscs - cms的冷冻保存培养基配方，尽管它们的价格和成分不同，但解冻后的回收率和细胞功能大致相似。©2019作者。基本方案1:解离和低温保存hpscs - cms基本方案2:解冻和低温冷冻的hpscs - cms恢复
本文章由计算机程序翻译，如有差异，请以英文原文为准。
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