Mu Junsheng, Gao Zhen, Bo Ping, You Bin
Department of Cardiac Surgery, Beijing Anzhen Hospital, Capital Medical University, Beijing Institute of Heart Lung and Blood Vessel Diseases, Beijing, China.
The Third Affiliated Hospital of XinXiang Medical University, XinXiang, China.
Biomed Mater Eng. 2025 Jan;36(1):34-42. doi: 10.3233/BME-240101.
Myocardial infarction leads to myocardial necrosis, and cardiomyocytes are non-renewable. Fatty acid-containing cardiomyocyte maturation medium promotes maturation of stem cell-derived cardiomyocytes.
To study the effect palmitic acid on maturation of cardiomyocytes derived from human embryonic stem cells (hESCs) to optimize differentiation for potential treatment of myocardial infarction by hESCs.
hESCs were differentiated into cardiomyocytes using standard chemically defined medium 3 (CDM3). Up to day 20 of differentiation, 200 Mm palmitic acid were added, and then the culture was continued for another 8 days to mimic the environment in which human cardiomyocytes mainly use fatty acids as the main energy source. Light microscopy, transmission electron microscopy, immunofluorescence, reverse transcription-polymerase chain reaction, and cellular ATP assays, were carried out to analyze the expression of relevant cardiomyocyte-related genes, cell morphology, metabolism levels, and other indicators cardiomyocyte maturity.
Cardiomyocytes derived from hESCs under exogenous palmitic acid had an elongated pike shape and a more regular arrangement. Sarcomere stripes were clear, and the cells color was clearly visible. The cell perimeter and elongation rate were also increased. Myogenic fibers were abundant, myofibrillar z-lines were regularly, the numbers of mitochondria and mitochondrial cristae were higher, more myofilaments were observed, and the structure of round-like discs was occasionally seen. Expression of mature cardiomyocyte-associated genes TNNT2, MYL2 and MYH6, and cardiomyocyte-associated genes KCNJ4, RYR2,and PPARα, was upregulated ( < 0.05). Expression of MYH7, MYL7, KCND2, KCND3, GJA1 and TNNI1 genes was unaffected ( > 0.05). Expression of mature cardiomyocyte-associated sarcomere protein MYL2 was significantly increased ( < 0.05), MYH7 protein expression was unaffected ( > 0.05). hESC-derived cardiomyocytes exposed to exogenous palmitic acid produced more ATP per unit time ( < 0.05).
Exogenous palmitic acid induced more mature hESC-CMs in terms of the cellular architecture, expression of cardiomyocyte maturation genes adnprotein, and metabolism.
心肌梗死会导致心肌坏死，而心肌细胞不可再生。含脂肪酸的心肌细胞成熟培养基可促进干细胞衍生心肌细胞的成熟。
研究棕榈酸对人胚胎干细胞（hESCs）来源的心肌细胞成熟的影响，以优化分化过程，为hESCs潜在治疗心肌梗死提供依据。
使用标准化学成分明确培养基3（CDM3）将hESCs分化为心肌细胞。在分化的第20天添加200μM棕榈酸，然后继续培养8天，以模拟人类心肌细胞主要以脂肪酸为主要能量来源的环境。通过光学显微镜、透射电子显微镜、免疫荧光、逆转录-聚合酶链反应和细胞ATP测定，分析相关心肌细胞相关基因的表达、细胞形态、代谢水平和其他心肌细胞成熟指标。
外源性棕榈酸作用下hESCs来源的心肌细胞呈细长梭形，排列更规则。肌节条纹清晰，细胞颜色明显可见。细胞周长和伸长率也增加。肌原纤维丰富，肌原纤维z线规则，线粒体和线粒体嵴数量更多，观察到更多肌丝，偶尔可见类圆形盘状结构。成熟心肌细胞相关基因TNNT2、MYL2和MYH6以及心肌细胞相关基因KCNJ4、RYR2和PPARα的表达上调（P<0.05）。MYH7、MYL7、KCND2、KCND3、GJA1和TNNI1基因的表达未受影响（P>0.05）。成熟心肌细胞相关肌节蛋白MYL2的表达显著增加（P<0.05），MYH7蛋白表达未受影响（P>0.05）。暴露于外源性棕榈酸的hESCs来源的心肌细胞每单位时间产生更多的ATP（P<0.05）。
外源性棕榈酸在细胞结构、心肌细胞成熟基因和蛋白质表达以及代谢方面诱导hESC-CMs更成熟。
