Negligible-Cost and Weekend-Free Chemically Defined Human iPSC Culture
Negligible-Cost and Weekend-Free Chemically Defined Human iPSC Culture
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DOI：
10.1016/j.stemcr.2019.12.007
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发表时间：
2020-02-11
影响因子：
5.9
通讯作者：
Burridge, Paul W.
中科院分区：
文献类型：
--
作者：
Kuo, Hui-Hsuan;Gao, Xiaozhi;Burridge, Paul W.
人诱导多能干细胞（hiPSC）培养已经成为常规，但多能细胞培养基的成本、频繁的培养基更换和分化的再现性仍然受到限制。在这里，我们描述了作为培养基成分和浓度的彻底优化的结果的hiPSC培养基（B8）的配制，确定了每种组分对多能状态和细胞增殖的必要性和相对贡献。B8中的试剂仅占商业培养基成本的3%，主要通过实验室内生成三种E。大肠杆菌表达的、密码子优化的重组蛋白：成纤维细胞生长因子2、转化生长因子B3和神经调节蛋白1。我们证明了B8中34个hiPSC系的衍生和培养以及多能性的长期维持（超过100代）。该配方还允许无周末喂养时间表，而不牺牲分化能力。
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 b3, 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.