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

Davi M. Lyra‐Leite, Hananeh Fonoudi, Mennat Gharib, Paul W. Burridge

2021STAR Protocols17 citationsDOIOpen Access PDF

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).

Topics & Concepts

TroubleshootingMatrigelProtocol (science)Induced pluripotent stem cellComputer scienceHuman Induced Pluripotent Stem CellsChemistryBiologyCell cultureOperating systemBiochemistryMedicineGeneticsEmbryonic stem cellPathologyGeneAlternative medicinePluripotent Stem Cells Research3D Printing in Biomedical ResearchCRISPR and Genetic Engineering