Department of Cardiology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan; Department of Organ Fabrication, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan.
Department of Cardiology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan.
Stem Cell Reports. 2017 Nov 14;9(5):1406-1414. doi: 10.1016/j.stemcr.2017.08.025. Epub 2017 Oct 5.
Cardiac regenerative therapies utilizing human induced pluripotent stem cells (hiPSCs) are hampered by ineffective large-scale culture. hiPSCs were cultured in multilayer culture plates (CPs) with active gas ventilation (AGV), resulting in stable proliferation and pluripotency. Seeding of 1 × 10 hiPSCs per layer yielded 7.2 × 10 hiPSCs in 4-layer CPs and 1.7 × 10 hiPSCs in 10-layer CPs with pluripotency. hiPSCs were sequentially differentiated into cardiomyocytes (CMs) in a two-dimensional (2D) differentiation protocol. The efficiency of cardiac differentiation using 10-layer CPs with AGV was 66%-87%. Approximately 6.2-7.0 × 10 cells (4-layer) and 1.5-2.8 × 10 cells (10-layer) were obtained with AGV. After metabolic purification with glucose- and glutamine-depleted and lactate-supplemented media, a massive amount of purified CMs was prepared. Here, we present a scalable 2D culture system using multilayer CPs with AGV for hiPSC-derived CMs, which will facilitate clinical applications for severe heart failure in the near future.
利用人诱导多能干细胞（hiPSC）的心脏再生疗法受到大规模培养效果不佳的阻碍。hiPSC 在具有主动气体通气（AGV）的多层培养板（CP）中进行培养，从而实现稳定增殖和多能性。每层层播种 1×10 的 hiPSC 可在 4 层 CP 中产生 7.2×10 的 hiPSC，在 10 层 CP 中产生 1.7×10 的 hiPSC，且具有多能性。hiPSC 按照二维（2D）分化方案依次分化为心肌细胞（CM）。使用具有 AGV 的 10 层 CP 进行心脏分化的效率为 66%-87%。使用 AGV 可获得约 6.2-7.0×10 个细胞（4 层）和 1.5-2.8×10 个细胞（10 层）。在使用葡萄糖和谷氨酰胺耗尽并补充乳酸盐的代谢纯化后，可制备大量纯化的 CM。在此，我们提出了一种使用具有 AGV 的多层 CP 的可扩展 2D 培养系统，用于 hiPSC 衍生的 CM，这将有助于在不久的将来为严重心力衰竭的临床应用提供便利。
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