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一种用于人类诱导多能干细胞经济高效且无需周末培养的更新方案。

An updated protocol for the cost-effective and weekend-free culture of human induced pluripotent stem cells.

作者信息

Lyra-Leite Davi Marco, Fonoudi Hananeh, Gharib Mennat, Burridge Paul W

机构信息

Department of Pharmacology, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA.

Center for Pharmacogenomics, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA.

出版信息

STAR Protoc. 2021 Feb 3;2(1):100213. doi: 10.1016/j.xpro.2020.100213. eCollection 2021 Mar 19.

DOI:10.1016/j.xpro.2020.100213
PMID:33786455
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7988236/
Abstract

The protocol provided here describes methodologies for making a highly cost-effective, chemically defined medium for culturing hiPSCs we call B8 medium. The typical cost of B8 medium is US$10 per liter, which with modifications included here is more affordable than standard media. We provide simple protocols for making B8 supplement aliquots, making the basal media DMEM/F12, Matrigel-coated plates, thawing, passaging, culturing, and cryopreserving hiPSCs. We show typical differentiation results and provide a comprehensive troubleshooting guide. For complete details on the use and execution of this protocol, please refer to Kuo et al. (2020).

摘要

此处提供的方案描述了一种用于培养人诱导多能干细胞（hiPSC）的高性价比、化学成分明确的培养基（我们称之为B8培养基）的制备方法。B8培养基的典型成本为每升10美元，经过此处所述的改进后，比标准培养基更经济实惠。我们提供了制备B8补充剂等分试样、制备基础培养基DMEM/F12、基质胶包被板、解冻、传代、培养和冻存hiPSC的简单方案。我们展示了典型的分化结果，并提供了一份全面的故障排除指南。有关本方案使用和执行的完整详细信息，请参考Kuo等人（2020年）的文献。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8349/7988236/d04c1091aace/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8349/7988236/db4e090b7098/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8349/7988236/ba5ba42e7d6a/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8349/7988236/7dd8f974ee96/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8349/7988236/ea0d2d6453c0/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8349/7988236/c9ee071b6ee2/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8349/7988236/d04c1091aace/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8349/7988236/db4e090b7098/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8349/7988236/ba5ba42e7d6a/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8349/7988236/7dd8f974ee96/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8349/7988236/ea0d2d6453c0/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8349/7988236/c9ee071b6ee2/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8349/7988236/d04c1091aace/gr5.jpg

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