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

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

doi:10.1016/j.xpro.2020.100213

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

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

Authors

Davi Marco Lyra-Leite^{1

2}, Hananeh Fonoudi^{1

2}, Mennat Gharib^{1

2}, Paul W Burridge^{1

2}

Affiliations

¹ Department of Pharmacology, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA.
² Center for Pharmacogenomics, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA.

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).

Keywords: Cell Culture; Stem Cells.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Cell Culture Techniques*
Cell Differentiation*
Culture Media / chemistry
Culture Media / pharmacology
Humans
Induced Pluripotent Stem Cells / cytology*
Induced Pluripotent Stem Cells / metabolism*

Substances

Culture Media

Grants and funding

R01 CA220002/CA/NCI NIH HHS/United States