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.

人类诱导多能干细胞 (hiPSC) 培养已成为常规，但多能细胞培养基的成本、频繁的培养基更换以及分化的可重复性仍然受到限制。在这里，我们描述了 hiPSC 培养基 (B8) 的配方，这是对培养基成分和浓度进行彻底优化的结果，确定了每种成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%，这主要是通过在实验室中生成三种大肠杆菌表达、密码子优化的重组蛋白而实现的：成纤维细胞生长因子 2、转化生长因子 β3 和神经调节蛋白1. 我们展示了 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计划，而不会牺牲分化能力。