Promotion of maturation in CDM3-induced embryonic stem cell-derived cardiomyocytes by palmitic acid.

IF 1 4区医学 Q4 ENGINEERING, BIOMEDICAL

Bio-medical materials and engineering Pub Date : 2024-09-26 DOI:10.3233/BME-240101

Junsheng Mu, Zhen Gao, Ping Bo, Bin You

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引用次数: 0

Abstract

Background: Myocardial infarction leads to myocardial necrosis, and cardiomyocytes are non-renewable. Fatty acid-containing cardiomyocyte maturation medium promotes maturation of stem cell-derived cardiomyocytes.

Objective: 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.

Methods: 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.

Results: 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 (p < 0.05). Expression of MYH7, MYL7, KCND2, KCND3, GJA1 and TNNI1 genes was unaffected (p > 0.05). Expression of mature cardiomyocyte-associated sarcomere protein MYL2 was significantly increased (p < 0.05), MYH7 protein expression was unaffected (p > 0.05). hESC-derived cardiomyocytes exposed to exogenous palmitic acid produced more ATP per unit time (p < 0.05).

Conclusion: Exogenous palmitic acid induced more mature hESC-CMs in terms of the cellular architecture, expression of cardiomyocyte maturation genes adnprotein, and metabolism.

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棕榈酸促进 CDM3 诱导的胚胎干细胞衍生心肌细胞的成熟。

背景：心肌梗死导致心肌坏死，而心肌细胞是不可再生的。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：研究棕榈酸对人类胚胎干细胞（hESCs）衍生的心肌细胞成熟的影响，以优化分化，从而利用 hESCs 治疗心肌梗死。方法：使用标准化学定义培养基 3（CDM3）将 hESCs 分化成心肌细胞。方法：使用标准化学定义培养基 3（CDM3）将 hESCs 分化成心肌细胞，在分化的第 20 天，加入 200 Mm 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、反转录聚合酶链反应和细胞ATP测定等方法，分析心肌细胞相关基因的表达、细胞形态、代谢水平等心肌细胞成熟度指标：结果：在外源性棕榈酸的作用下，由 hESCs 培育出的心肌细胞呈拉长的梭形，排列更加规则。肌节条纹清晰，细胞颜色明显。细胞周长和伸长率也有所增加。肌原纤维丰富，肌纤维 Z 线规则，线粒体和线粒体嵴数量增多，肌丝增多，偶见圆盘状结构。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 表达上调（p 0.05）。暴露于外源棕榈酸的 hESC 衍生心肌细胞在单位时间内产生更多的 ATP（p 结论：外源棕榈酸诱导心肌细胞产生更多的 ATP（p 结论：外源棕榈酸诱导心肌细胞产生更多的 ATP（p 结论：外源棕榈酸诱导心肌细胞产生更多的 ATP（p 结论：外源棕榈酸诱导心肌细胞产生更多的 ATP（p 结论）：外源性棕榈酸可诱导 hESC-CMs 在细胞结构、心肌细胞成熟基因 adnprotein 的表达和新陈代谢方面更加成熟。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Bio-medical materials and engineering 工程技术-材料科学：生物材料

CiteScore

1.80

自引率

0.00%

发文量

审稿时长

6 months

期刊介绍： The aim of Bio-Medical Materials and Engineering is to promote the welfare of humans and to help them keep healthy. This international journal is an interdisciplinary journal that publishes original research papers, review articles and brief notes on materials and engineering for biological and medical systems. Articles in this peer-reviewed journal cover a wide range of topics, including, but not limited to: Engineering as applied to improving diagnosis, therapy, and prevention of disease and injury, and better substitutes for damaged or disabled human organs; Studies of biomaterial interactions with the human body, bio-compatibility, interfacial and interaction problems; Biomechanical behavior under biological and/or medical conditions; Mechanical and biological properties of membrane biomaterials; Cellular and tissue engineering, physiological, biophysical, biochemical bioengineering aspects; Implant failure fields and degradation of implants. Biomimetics engineering and materials including system analysis as supporter for aged people and as rehabilitation; Bioengineering and materials technology as applied to the decontamination against environmental problems; Biosensors, bioreactors, bioprocess instrumentation and control system; Application to food engineering; Standardization problems on biomaterials and related products; Assessment of reliability and safety of biomedical materials and man-machine systems; and Product liability of biomaterials and related products.