Generating a Cost-Effective, Weekend-Free Chemically Defined Human Induced Pluripotent Stem Cell (hiPSC) Culture Medium

Q2 Biochemistry, Genetics and Molecular Biology

Current Protocols in Stem Cell Biology Pub Date : 2020-05-28 DOI:10.1002/cpsc.110

Hananeh Fonoudi, Davi M. Lyra-Leite, Hoor A. Javed, Paul W. Burridge

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引用次数: 1

Abstract

We have previously developed a cost-effective chemically defined medium formula for weekend-free culture of human induced pluripotent stem cells (hiPSCs), costing ∼3% of the price of commercial medium. This medium, which we termed B8, is specifically optimized for robust and fast growth of hiPSCs and for a weekend-free medium change regimen. We demonstrated that this medium is suitable for reprogramming of somatic cells into hiPSCs and for differentiation into a variety of lineages. Here, we provide a protocol for simple generation of the most cost-effective variant of this medium, along with a protocol for making Matrigel-coated plates and culturing, passaging, cryopreserving, and thawing hiPSCs. © 2020 Wiley Periodicals LLC.

Basic Protocol 1: Preparation of a highly optimized, robust, and cost-effective human induced pluripotent stem cell culture medium

Basic Protocol 2: Weekend-free maintenance and passaging of human induced pluripotent stem cells in B8 medium

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制备一种低成本、无周末化学定义的人诱导多能干细胞(hiPSC)培养基

我们之前开发了一种具有成本效益的化学定义培养基配方，用于人诱导多能干细胞(hiPSCs)的周末无培养，成本为商业培养基价格的3%。我们将这种培养基命名为B8，专门针对hipsc的强劲快速生长和周末无培养基更换方案进行了优化。我们证明了这种培养基适合将体细胞重编程为hipsc并分化为各种谱系。在这里，我们提供了一种最具成本效益的培养基变体的简单生成方案，以及制作matrigel涂层板和培养，传代，冷冻保存和解冻hipsc的方案。©2020 Wiley期刊有限公司基本方案1:制备高度优化、稳健且具有成本效益的人诱导多能干细胞培养基基本方案2:在B8培养基中无周末维持和传代人诱导多能干细胞

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

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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.