Cell Banking of hiPSCs: A Practical Guide to Cryopreservation and Quality Control in Basic Research

Q2 Biochemistry, Genetics and Molecular Biology Current Protocols in Stem Cell Biology Pub Date : 2020-09-21 DOI:10.1002/cpsc.127

Aya Shibamiya, Elisabeth Schulze, Dana Krauß, Christa Augustin, Marina Reinsch, Mirja Loreen Schulze, Simone Steuck, Giulia Mearini, Ingra Mannhardt, Thomas Schulze, Birgit Klampe, Tessa Werner, Umber Saleem, Anika Knaust, Sandra D. Laufer, Christiane Neuber, Marta Lemme, Charlotta Sophie Behrens, Malte Loos, Florian Weinberger, Sigrid Fuchs, Thomas Eschenhagen, Arne Hansen, Bärbel Maria Ulmer

{"title":"Cell Banking of hiPSCs: A Practical Guide to Cryopreservation and Quality Control in Basic Research","authors":"Aya Shibamiya, Elisabeth Schulze, Dana Krauß, Christa Augustin, Marina Reinsch, Mirja Loreen Schulze, Simone Steuck, Giulia Mearini, Ingra Mannhardt, Thomas Schulze, Birgit Klampe, Tessa Werner, Umber Saleem, Anika Knaust, Sandra D. Laufer, Christiane Neuber, Marta Lemme, Charlotta Sophie Behrens, Malte Loos, Florian Weinberger, Sigrid Fuchs, Thomas Eschenhagen, Arne Hansen, Bärbel Maria Ulmer","doi":"10.1002/cpsc.127","DOIUrl":null,"url":null,"abstract":"The reproducibility of stem cell research relies on the constant availability of quality-controlled cells. As the quality of human induced pluripotent stem cells (hiPSCs) can deteriorate in the course of a few passages, cell banking is key to achieve consistent results and low batch-to-batch variation. Here, we provide a cost-efficient route to generate master and working cell banks for basic research projects. In addition, we describe minimal protocols for quality assurance including tests for sterility, viability, pluripotency, and genetic integrity. © 2020 The Authors.Basic Protocol 1: Expansion of hiPSCsBasic Protocol 2: Cell banking of hiPSCsSupport Protocol 1: Pluripotency assessment by flow cytometrySupport Protocol 2: Thawing control: Viability and sterilitySupport Protocol 3: Potency, viral clearance, and pluripotency: Spontaneous differentiation and qRT-PCRSupport Protocol 4: Identity: Short tandem repeat analysis","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.127","citationCount":"9","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Stem Cell Biology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpsc.127","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}

引用次数: 9

Abstract

The reproducibility of stem cell research relies on the constant availability of quality-controlled cells. As the quality of human induced pluripotent stem cells (hiPSCs) can deteriorate in the course of a few passages, cell banking is key to achieve consistent results and low batch-to-batch variation. Here, we provide a cost-efficient route to generate master and working cell banks for basic research projects. In addition, we describe minimal protocols for quality assurance including tests for sterility, viability, pluripotency, and genetic integrity. © 2020 The Authors.

Basic Protocol 1: Expansion of hiPSCs

Basic Protocol 2: Cell banking of hiPSCs

Support Protocol 1: Pluripotency assessment by flow cytometry

Support Protocol 2: Thawing control: Viability and sterility

Support Protocol 3: Potency, viral clearance, and pluripotency: Spontaneous differentiation and qRT-PCR

Support Protocol 4: Identity: Short tandem repeat analysis

Abstract Image

查看原文

微信好友朋友圈 QQ好友复制链接

本刊更多论文

hiPSCs的细胞库:基础研究中低温保存和质量控制的实用指南

干细胞研究的可重复性依赖于质量控制细胞的持续可用性。由于人类诱导多能干细胞(hiPSCs)的质量在几代传代过程中可能会恶化，细胞库是实现一致结果和低批次差异的关键。在这里，我们为基础研究项目提供了一种具有成本效益的途径来生成主细胞库和工作细胞库。此外，我们还描述了质量保证的最小方案，包括无菌性、活力、多能性和遗传完整性测试。©2020作者。基本方案1:hipsscs的扩增基本方案2:hipsscs的细胞库支持方案1:流式细胞术多能性评估支持方案2:解冻控制:活力和无菌支持方案3:效力，病毒清除和多能性:自发分化和qrt - pcr支持方案4:身份:短串联重复分析

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文去求助

来源期刊

Current Protocols in Stem Cell Biology Biochemistry, Genetics and Molecular Biology-Cell Biology

自引率

0.00%

发文量

期刊介绍： Published in affiliation with the International Society for Stem Cell Research (ISSCR), Current Protocols in Stem Cell Biology (CPSC) covers the most fundamental protocols and methods in the rapidly growing field of stem cell biology. Updated monthly, CPSC will constantly evolve with thelatest developments and breakthroughs in the field. Drawing on the expertise of leading researchers from around the world, Current Protocols in Stem Cell Biology includes methods and insights that will enhance the progress of global research.