Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University (TWIns), 8-1 Kawada-cho, Shinjuku-ku, Tokyo, 162-8666, Japan.
Sci Rep. 2024 May 15;14(1):11081. doi: 10.1038/s41598-024-60765-5.
Despite progress in generating cardiomyocytes from pluripotent stem cells, these populations often include non-contractile cells, necessitating cardiomyocyte selection for experimental purpose. This study explores a novel cardiomyocyte enrichment mechanism: low-adhesion culture selection. The cardiac cells derived from human induced pluripotent stem cells were subjected to a coating-free low-adhesion culture using bovine serum albumin and high molecular weight dextran sulfate. This approach effectively increased the population of cardiac troponin T-positive cardiomyocytes. Similar results were obtained with commercially available low-adhesion culture dishes. Subsequently, we accessed the practicality of selection of cardiomyocytes using this phenomenon by comparing it with established methods such as glucose-free culture and selection based on puromycin resistance genes. The cardiomyocytes enriched through low-adhesion culture selection maintained autonomous pulsation and responsiveness to beta-stimuli. Moreover, no significant differences were observed in the expression of genes related to subtype commitment and maturation when compared to other selection methods. In conclusion, cardiomyocytes derived from pluripotent stem cells were more low-adhesion culture resistant than their accompanying non-contractile cells, and low-adhesion culture is an alternative method for selection of pluripotent stem cell-derived cardiomyocytes.
尽管在从多能干细胞中生成心肌细胞方面取得了进展,但这些细胞群体通常包含非收缩性细胞,因此需要对心肌细胞进行选择,以用于实验目的。本研究探索了一种新颖的心肌细胞富集机制:低黏附培养选择。将源自人诱导多能干细胞的心脏细胞用牛血清白蛋白和高分子量硫酸葡聚糖进行无涂层的低黏附培养。这种方法有效地增加了心肌肌钙蛋白 T 阳性心肌细胞的数量。使用市售的低黏附培养皿也获得了类似的结果。随后,我们通过将其与葡萄糖饥饿培养和基于嘌呤霉素抗性基因的选择等已建立的方法进行比较,评估了使用这种现象进行心肌细胞选择的实用性。通过低黏附培养选择富集的心肌细胞保持自主搏动和对β刺激的反应性。此外,与其他选择方法相比,在与亚型决定和成熟相关的基因表达方面未观察到显著差异。总之,与伴随的非收缩性细胞相比,多能干细胞来源的心肌细胞对低黏附培养的抗性更强,低黏附培养是一种用于选择多能干细胞衍生的心肌细胞的替代方法。