Efficient Large-Scale 2D Culture System for Human Induced Pluripotent Stem Cells and Differentiated Cardiomyocytes.
Efficient Large-Scale 2D Culture System for Human Induced Pluripotent Stem Cells and Differentiated Cardiomyocytes.
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DOI：
10.1016/j.stemcr.2017.08.025
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发表时间：
2017-11-14
影响因子：
5.9
通讯作者：
Fukuda K
中科院分区：
文献类型：
--
作者：
Tohyama S;Fujita J;Fujita C;Yamaguchi M;Kanaami S;Ohno R;Sakamoto K;Kodama M;Kurokawa J;Kanazawa H;Seki T;Kishino Y;Okada M;Nakajima K;Tanosaki S;Someya S;Hirano A;Kawaguchi S;Kobayashi E;Fukuda K
利用人诱导多能干细胞（hiPSC）的心脏再生疗法受到无效的大规模培养的阻碍。hiPSC在具有主动气体通气（AGV）的多层培养板（CP）中培养，导致稳定的增殖和多能性。每层接种1 × 106个hiPSC，在4层CP中产生7.2 × 108个hiPSC，在10层CP中产生1.7 × 109个具有多能性的hiPSC。hiPSC在二维（2D）分化方案中依次分化成心肌细胞（CM）。使用10层CP和AGV进行心脏分化的效率为66%-87%。用AGV获得约6.2-7.0 × 108个细胞（4层）和1.5-2.8 × 109个细胞（10层）。在用葡萄糖和谷氨酰胺耗尽和乳酸补充的培养基进行代谢纯化后，制备了大量纯化的CM。在这里，我们提出了一个可扩展的二维培养系统，使用多层CP与AGV的hiPSC衍生的CM，这将有助于在不久的将来严重心力衰竭的临床应用。通过具有AGV的多层培养板高效大规模生产hiPSC使用具有AGV的大规模2D培养系统高效大规模生产hiPSC-CM通过代谢选择大规模生产纯hiPSC-CM在这篇文章中，Fujita及其同事表明，人类诱导多能干细胞（hiPSC）和hiPSC衍生的心肌细胞由于培养基中稳定的CO2和O2浓度，使用具有主动气体通风系统的多层培养板稳定且有效地生产hiPSC-CM。大量获得的hiPSC-CM在葡萄糖和谷氨酰胺耗尽和乳酸盐补充条件下代谢纯化。
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 × 106 hiPSCs per layer yielded 7.2 × 108 hiPSCs in 4-layer CPs and 1.7 × 109 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 × 108 cells (4-layer) and 1.5–2.8 × 109 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. Efficient mass production of hiPSCs by multilayer culture plates with AGV Efficient mass production of hiPSC-CMs using a massive 2D culture system with AGV Mass production of pure hiPSC-CMs via metabolic selection In this article, Fujita and colleagues show that human induced pluripotent stem cells (hiPSCs) and hiPSC-derived cardiomyocytes (hiPSC-CMs) are stably and efficiently produced using multilayer culture plates with an active gas ventilation system because of stable CO2 and O2 concentration in culture media. Large numbers of obtained hiPSC-CMs are metabolically purified under glucose- and glutamine-depleted and lactate-supplemented conditions.
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影响因子：
3.7
作者：
Badenes SM;Fernandes TG;Cordeiro CS;Boucher S;Kuninger D;Vemuri MC;Diogo MM;Cabral JM
通讯作者：
Cabral JM
影响因子：
20.1
作者：
Zhang J;Wilson GF;Soerens AG;Koonce CH;Yu J;Palecek SP;Thomson JA;Kamp TJ
通讯作者：
Kamp TJ
影响因子：
3
作者：
Lecina, Marti;Ting, Sherwin;Oh, Steve
通讯作者：
Oh, Steve
影响因子：
6
作者：
Silva, Marta M.;Rodrigues, Ana F.;Alves, Paula M.
通讯作者：
Alves, Paula M.
影响因子：
3.7
作者：
Schulz TC;Young HY;Agulnick AD;Babin MJ;Baetge EE;Bang AG;Bhoumik A;Cepa I;Cesario RM;Haakmeester C;Kadoya K;Kelly JR;Kerr J;Martinson LA;McLean AB;Moorman MA;Payne JK;Richardson M;Ross KG;Sherrer ES;Song X;Wilson AZ;Brandon EP;Green CE;Kroon EJ;Kelly OG;D'Amour KA;Robins AJ
通讯作者：
Robins AJ