Negligible-Cost and Weekend-Free Chemically Defined Human iPSC Culture
| العنوان: | Negligible-Cost and Weekend-Free Chemically Defined Human iPSC Culture |
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| المؤلفون: | Tarek Magdy, Hui Hsuan Kuo, Alfred L. George, K. Ashley Fetterman, Paul W. Burridge, Mariam Jouni, Jean Marc DeKeyser, Marisol Romero-Tejeda, Emily Pinheiro, Hananeh Fonoudi, Michael V. Orman, Malorie Blancard, Conrad L. Epting, Carly J. Weddle, Xiaozhi Gao |
| المصدر: | Stem Cell Reports, Vol 14, Iss 2, Pp 256-270 (2020) Stem Cell Reports |
| بيانات النشر: | Cold Spring Harbor Laboratory, 2019. |
| سنة النشر: | 2019 |
| مصطلحات موضوعية: | 0301 basic medicine, culture media, FGF2, Cell, Induced Pluripotent Stem Cells, Cell Culture Techniques, Fibroblast growth factor, Regenerative medicine, Biochemistry, Article, chemically defined, 03 medical and health sciences, 0302 clinical medicine, medicine, Genetics, Humans, Neuregulin 1, pluripotent state, Induced pluripotent stem cell, weekend-free, lcsh:QH301-705.5, Cells, Cultured, 030304 developmental biology, Cell Proliferation, 0303 health sciences, lcsh:R5-920, biology, Chemistry, Cell growth, human induced pluripotent stem cell, Cell Differentiation, Cell Biology, differentiation, Cell biology, Open source, medicine.anatomical_structure, 030104 developmental biology, lcsh:Biology (General), biology.protein, Biological Assay, lcsh:Medicine (General), 030217 neurology & neurosurgery, Developmental Biology, Transforming growth factor |
| الوصف: | Summary Human induced pluripotent stem cell (hiPSC) culture has become routine, yet the cost of pluripotent cell media, frequent medium changes, and the reproducibility of differentiation have remained restrictive. Here, we describe the formulation of a hiPSC culture medium (B8) as a result of the exhaustive optimization of medium constituents and concentrations, establishing the necessity and relative contributions of each component to the pluripotent state and cell proliferation. The reagents in B8 represent only 3% of the costs of commercial media, made possible primarily by the in-lab generation of three E. coli-expressed, codon-optimized recombinant proteins: fibroblast growth factor 2, transforming growth factor β3, and neuregulin 1. We demonstrate the derivation and culture of 34 hiPSC lines in B8 as well as the maintenance of pluripotency long term (over 100 passages). This formula also allows a weekend-free feeding schedule without sacrificing capacity for differentiation. Highlights • The B8 hiPSC culture medium formula is a result of exhaustive optimization • The reagents in B8 represent only 3% of the costs of commercial media • B8 is suitable for the derivation and culture of hiPSC lines long term (>100 passages) • This formula allows weekend-free feeding without sacrificing differentiation capacity In this article, Burridge and colleagues describe the formulation of a hiPSC culture medium called B8 as a result of exhaustive optimization. The reagents in B8 represent only 3% of the costs of commercial media. B8 is suitable for both derivation and long-term culture of hiPSCs and allows a weekend-free feeding schedule without sacrificing capacity for differentiation. |
| اللغة: | English |
| DOI: | 10.1101/685503 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::348e5ed3894e738e36d8f759e0656df3Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....348e5ed3894e738e36d8f759e0656df3 |
| قاعدة البيانات: | OpenAIRE |
| DOI: | 10.1101/685503 |
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