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单层培养人心细胞的耗氧量。

Oxygen consumption of human heart cells in monolayer culture.

机构信息

Department of Pediatrics, Tokai University School of Medicine, 143 Shimokasuya, Isehara, Kanagawa 259-1193, Japan.

Institute for Nanoscience and Nanotechnology, Waseda University, 2-2 Wakamatsu-cho, Shinjuku-ku, Tokyo 162-8480, Japan.

出版信息

Biochem Biophys Res Commun. 2014 Sep 26;452(3):834-9. doi: 10.1016/j.bbrc.2014.09.018. Epub 2014 Sep 16.

Abstract

Tissue engineering in cardiovascular regenerative therapy requires the development of an efficient oxygen supply system for cell cultures. However, there are few studies which have examined human cardiomyocytes in terms of oxygen consumption and metabolism in culture. We developed an oxygen measurement system equipped with an oxygen microelectrode sensor and estimated the oxygen consumption rates (OCRs) by using the oxygen concentration profiles in culture medium. The heart is largely made up of cardiomyocytes, cardiac fibroblasts, and cardiac endothelial cells. Therefore, we measured the oxygen consumption of human induced pluripotent stem cell derived cardiomyocytes (hiPSC-CMs), cardiac fibroblasts, human cardiac microvascular endothelial cell and aortic smooth muscle cells. Then we made correlations with their metabolisms. In hiPSC-CMs, the value of the OCR was 0.71±0.38pmol/h/cell, whereas the glucose consumption rate and lactate production rate were 0.77±0.32pmol/h/cell and 1.61±0.70pmol/h/cell, respectively. These values differed significantly from those of the other cells in human heart. The metabolism of the cells that constitute human heart showed the molar ratio of lactate production to glucose consumption (L/G ratio) that ranged between 1.97 and 2.2. Although the energy metabolism in adult heart in vivo is reported to be aerobic, our data demonstrated a dominance of anaerobic glycolysis in an in vitro environment. With our measuring system, we clearly showed the differences in the metabolism of cells between in vivo and in vitro monolayer culture. Our results regarding cell OCRs and metabolism may be useful for future tissue engineering of human heart.

摘要

组织工程学在心血管再生治疗中需要开发一种高效的细胞培养供氧系统。然而,关于培养中的人类心肌细胞的氧消耗和代谢,很少有研究进行过探讨。我们开发了一种配备氧微电极传感器的氧测量系统,并通过测量培养基中的氧浓度分布来估计氧消耗率(OCR)。心脏主要由心肌细胞、心脏成纤维细胞和心脏内皮细胞组成。因此,我们测量了人类诱导多能干细胞衍生的心肌细胞(hiPSC-CMs)、心脏成纤维细胞、人类心脏微血管内皮细胞和主动脉平滑肌细胞的氧消耗。然后,我们将其与代谢物进行了相关性分析。在 hiPSC-CMs 中,OCR 值为 0.71±0.38pmol/h/细胞,而葡萄糖消耗率和乳酸生成率分别为 0.77±0.32pmol/h/细胞和 1.61±0.70pmol/h/细胞。这些值与人心肌中的其他细胞有显著差异。构成人心肌的细胞代谢显示出乳酸生成与葡萄糖消耗的摩尔比(L/G 比)在 1.97 到 2.2 之间。尽管体内成年心脏的能量代谢被报道为有氧的,但我们的数据表明,在体外环境中,无氧糖酵解占主导地位。通过我们的测量系统,我们清楚地显示了细胞在体内和体外单层培养中的代谢差异。我们关于细胞 OCR 和代谢的结果可能对未来人心肌的组织工程有用。

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