Development of appropriate fatty acid formulations to raise the contractility of constructed myocardial tissues.
Development of appropriate fatty acid formulations to raise the contractility of constructed myocardial tissues.
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DOI：
10.1016/j.reth.2022.09.006
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发表时间：
2022-12
影响因子：
4.3
通讯作者：
Shimizu, Tatsuya
中科院分区：
文献类型：
--
作者：
Yoshida, Azumi;Sekine, Waki;Homma, Jun;Sekine, Hidekazu;Itoyama, Yu Yamasaki;Sasaki, Daisuke;Matsuura, Katsuhisa;Kobayashi, Eiji;Shimizu, Tatsuya
心脏病是世界范围内死亡的主要原因，近年来每年因心脏病死亡的人数有所增加。虽然心力衰竭通常通过药物治疗，但终末期疾病的最终治疗方法是心脏移植或人工心脏。然而，这些手术策略的使用受到血栓形成、排斥反应和供体短缺等问题的限制。再生疗法，如使用组织工程技术构建的培养细胞和组织的移植，作为心力衰竭的可能替代疗法受到极大关注。研究正在进行中，以潜在的临床应用的心肌细胞来源于人类诱导多能干细胞（hiPSC-CM）。然而，由于不成熟和对葡萄糖代谢的依赖，在先前培养条件下维持的心肌细胞的能量产生能力低于成年原代心肌细胞。因此，本研究的目的是比较培养的心肌细胞和体内的心脏细胞之间代谢的脂肪酸的类型，并研究向培养基中添加脂肪酸是否影响心肌细胞的能量产生。基于对大鼠原代心肌细胞（rat-CMs）脂肪酸消耗的分析开发了含脂肪酸的培养基，并通过表达腺苷三磷酸酶的rat-CMs的生物发光成像研究了该培养基对三磷酸腺苷（ATP）产生的影响。接下来，基于猪心和hiPSC-CM对脂肪酸利用的分析，进一步调整培养基的脂肪酸含量。进行耗氧分析以探索含脂肪酸的培养基是否诱导hiPSC-CM从厌氧代谢转换为需氧代谢。此外，评价了培养基对hiPSC-CM衍生组织产生的收缩力的影响。大鼠血清、人血清和猪血浆含有相似类型的脂肪酸（油酸、硬脂酸、亚油酸、棕榈酸和花生四烯酸）。大鼠-CM、hiPSC-CM和猪心之间消耗的脂肪酸类型也相似。向培养基中添加脂肪酸增加了表达腺苷三磷酸酶的大鼠-CM的生物发光（ATP水平的间接测量）、hiPSC-CM的氧消耗以及由hiPSC-CM构建的心脏组织产生的收缩力。hiPSC-CM代谢与大鼠-CM和猪心消耗脂肪酸类型相似的脂肪酸。此外，向培养基中添加这些脂肪酸增加了大鼠-CM和hiPSC-CM的能量产生，并增强了由hiPSC-CM产生的心肌组织的收缩性。这些发现表明，向培养基中添加脂肪酸通过β-氧化刺激心肌细胞产生有氧能量。由于在标准培养基中培养的心肌细胞主要依赖于无氧葡萄糖代谢并保持在未成熟状态，因此值得进一步研究以确定向培养基中添加脂肪酸是否会提高hiPSC-CM和由这些细胞构建的心脏组织的能量产生能力和成熟度。优化培养的心肌细胞的代谢（需要高能量产生来维持其收缩功能）可能会改善hiPSC-CM衍生组织的性质，使其能够更好地用于疾病建模、药物筛选和心力衰竭的再生治疗。
Heart disease is a major cause of mortality worldwide, and the annual number of deaths due to heart disease has increased in recent years. Although heart failure is usually managed with medicines, the ultimate treatment for end-stage disease is heart transplantation or an artificial heart. However, the use of these surgical strategies is limited by issues such as thrombosis, rejection and donor shortages. Regenerative therapies, such as the transplantation of cultured cells and tissues constructed using tissue engineering techniques, are receiving great attention as possible alternative treatments for heart failure. Research is ongoing into the potential clinical use of cardiomyocytes derived from human induced pluripotent stem cells (hiPSC-CMs). However, the energy-producing capacity of cardiomyocytes maintained under previous culture conditions is lower than that of adult primary cardiomyocytes due to immaturity and a reliance on glucose metabolism. Therefore, the aims of this study were to compare the types of fatty acids metabolized between cardiomyocytes in culture and heart cells in vivo and investigate whether the addition of fatty acids to the culture medium affected energy production by cardiomyocytes. A fatty acid-containing medium was developed based on an analysis of fatty acid consumption by rat primary cardiomyocytes (rat-CMs), and the effects of this medium on adenosine triphosphate (ATP) production were investigated through bioluminescence imaging of luciferase-expressing rat-CMs. Next, the fatty acid content of the medium was further adjusted based on analyses of fatty acid utilization by porcine hearts and hiPSC-CMs. Oxygen consumption analyses were performed to explore whether the fatty acid-containing medium induced hiPSC-CMs to switch from anaerobic metabolism to aerobic metabolism. Furthermore, the effects of the medium on contractile force generated by hiPSC-CM-derived tissue were evaluated. Rat serum, human serum and porcine plasma contained similar types of fatty acid (oleic acid, stearic acid, linoleic acid, palmitic acid and arachidonic acid). The types of fatty acid consumed were also similar between rat-CMs, hiPSC-CMs and porcine heart. The addition of fatty acids to the culture medium increased the bioluminescence of luciferase-expressing rat-CMs (an indirect measure of ATP level), oxygen consumption by hiPSC-CMs, and contractile force generated by cardiac tissues constructed from hiPSC-CMs. hiPSC-CMs metabolize similar types of fatty acid to those consumed by rat-CMs and porcine hearts. Furthermore, the addition of these fatty acids to the culture medium increased energy production by rat-CMs and hiPSC-CMs and enhanced the contractility of myocardial tissue generated from hiPSC-CMs. These findings suggest that the addition of fatty acids to the culture medium stimulates aerobic energy production by cardiomyocytes through β-oxidation. Since cardiomyocytes cultured in standard media rely primarily on anaerobic glucose metabolism and remain in an immature state, further research is merited to establish whether the addition of fatty acids to the culture medium would improve the energy-producing capacity and maturity of hiPSC-CMs and cardiac tissue constructed from these cells. It is possible that optimizing the metabolism of cultured cardiomyocytes, which require high energy production to sustain their contractile function, will improve the properties of hiPSC-CM-derived tissue, allowing it to be better utilized for disease modeling, drug screening and regenerative therapies for heart failure.
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影响因子：
16.6
作者：
Funakoshi S;Fernandes I;Mastikhina O;Wilkinson D;Tran T;Dhahri W;Mazine A;Yang D;Burnett B;Lee J;Protze S;Bader GD;Nunes SS;Laflamme M;Keller G
通讯作者：
Keller G
影响因子：
11
作者：
Lee SY;Ku HC;Kuo YH;Yang KC;Tu PC;Chiu HL;Su MJ
通讯作者：
Su MJ
影响因子：
7.2
作者：
Endo Y;Homma J;Sekine H;Matsuura K;Shimizu T;Niinami H
通讯作者：
Niinami H
影响因子：
5.2
作者：
Robertson, Claire;Tran, David D.;George, Steven C.
通讯作者：
George, Steven C.
影响因子：
3.3
作者：
Komae, Hyoe;Sekine, Hidekazu;Shimizu, Tatsuya
通讯作者：
Shimizu, Tatsuya