The design of a thermoresponsive surface for the continuous culture of human pluripotent stem cells
The design of a thermoresponsive surface for the continuous culture of human pluripotent stem cells
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DOI：
10.1016/j.biomaterials.2019.119411
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发表时间：
2019-11-01
期刊：
影响因子：
14
通讯作者：
Higuchi, Akon
中科院分区：
文献类型：
--
作者：
Sung, Tzu-Cheng;Yang, Jia-Sin;Higuchi, Akon
干细胞的培养通常是基于批量培养，这种方法既费力又昂贵。我们在热反应皿表面连续培养人多能干细胞（hPSCs），通过降低热反应皿温度至低于较低临界溶液温度仅30分钟，将hPSCs部分分离到同一热反应皿上。剩余细胞在新鲜培养基中继续培养，在交换培养基中收获离体干细胞。通过在热响应培养皿表面涂覆聚（n-异丙基丙烯酰胺-co-苯乙烯）和接枝聚（丙烯酸-co-苯乙烯）或重组玻璃连接蛋白，以维持hPSC结合位点的多能性，制备hPSC连续培养10个周期。在热反应培养皿表面连续培养10个周期后，分离的细胞表达多能性蛋白，在体外和体内均具有向三种胚层细胞分化的能力。此外，分离的细胞分化为特定的细胞系，如心肌细胞，效率高。
Commonly, stem cell culture is based on batch-type culture, which is laborious and expensive. We continuously cultured human pluripotent stem cells (hPSCs) on thermoresponsive dish surfaces, where hPSCs were partially detached on the same thermoresponsive dish by decreasing the temperature of the thermoresponsive dish to be below the lower critical solution temperature for only 30 min. Then, the remaining cells were continuously cultured in fresh culture medium, and the detached stem cells were harvested in the exchanged culture medium. hPSCs were continuously cultured for ten cycles on the thermoresponsive dish surface, which was prepared by coating the surface with poly(N-isopropylacrylamide-co-styrene) and oligovitronectin-grafted poly(acrylic acid co-styrene) or recombinant vitronectin for hPSC binding sites to maintain hPSC pluripotency. After ten cycles of continuous culture on the thermoresponsive dish surface, the detached cells expressed pluripotency proteins and had the ability to differentiate into cells derived from the three germ layers in vitro and in vivo. Furthermore, the detached cells differentiated into specific cell lineages, such as cardiomyocytes, with high efficiency.