생리적 배양 온도가 mTOR 신호전달 및 미토콘드리아 대사를 통해 닭 근육 줄기세포 증식에 미치는 영향 = Physiological Temperature Enhances Chicken Muscle Satellite Cell Proliferation Through mTOR Signaling and Mitochondrial Metabolism

다국어 초록 (Multilingual Abstract)

Chicken muscle satellite cells (CMSCs) are known to have both self-renewal ability and myogenic differentiation potential, and are therefore considered important for cultured meat research. Since the efficiency of cultured meat production largely depends on the proliferation of these cells, it is essential to identify conditions that can maximize their growth. Among the various factors, culture temperature plays a critical role. Although 37℃ is generally used as the standard culture temperature for most cell types, it does not fully reflect the physiological body temperature of chickens, which range from 41℃ to 42℃.
In this study, CMSCs were maintained at four temperatures (37, 39, 41, and 43℃) to identify the optimal condition for proliferation and then investigate the related metabolic changes at that temperature. Results from cell counting and CCK-8 assays showed that cells cultured at 41℃ exhibited a higher proliferation rate than those cultured at the other temperatures. Based on this result, metabolic analyses were mainly performed by comparing cells maintained at 37℃ and 41℃. Under 41℃ conditions mitochondrial function and ATP production were also increased, and qRT-PCR analysis revealed higher expression of genes related to the mTOR signaling pathway. ROS levels were elevated at 41℃, but the expression of SOD1 did not change significantly, catalase expression was reduced. Additionally, no apparent differences were found in apoptosis-related gene expression.
Overall, these results indicate that 41℃ is the most suitable temperature for promoting the proliferation and mitochondrial metabolism of CMSCs. This finding suggests that adjusting the culture temperature to the physiological level of chickens can improve the efficiency of CMSC expansion, which may contribute to the development of cultured meat production systems.

목차 (Table of Contents)

• CONTENTS
• LIST OF TABLES AND FIGURES
• ABBREVIATIONS AND SYMBOL
• ABSTRACT
• Ⅰ. INTRODUCTION
• Ⅱ. BACKGROUND AND LITERATURE REVIEW
• 2.1 Global Food Shortages and Increasing Demand for Protein
• 2.2 Cultured Meat as a Sustainable Protein Source
• 2.3 Physiological Characteristics and Regulatory Mechanisms of Muscle Satellite Cells
• 2.4 Temperature-dependent metabolic and proliferative characteristics of chicken muscle satellite cells
• Ⅲ. MATERIAL AND METHODS
• 3.1 Chicken muscle satellite cells isolation
• 3.2 Chicken muscle satellite cells culture
• 3.3 Cell viability analysis
• 3.4 Cell proliferation analysis
• 3.5 Immunofluorescence staining
• 3.6 Cell cycle analysis
• 3.7 Intracellular ROS analysis
• 3.8 Analysis of protein expression by Western blot
• 3.9 Analysis of gene expression by qRT-PCR
• 3.10 Measurement of Oxygen Consumption Rate
• 3.11 Statistical analysis
• Ⅳ. RESULTS
• 4.1 Effects of Temperature Treatment on Proliferation and Viability
• 4.2 Effects of Temperature Treatment on Myogenic Transcription Factor Expression
• 4.3 Effects of Temperature Treatment on the Cell Cycle
• 4.4 Effects of Temperature Treatment on Intracellular ROS, Antioxidant Response, and Apoptosis
• 4.5 Effects of Temperature Treatment on Mitochondrial Metabolism
• Ⅴ. DISCUSSION
• Ⅵ. CONCLUSION
• Ⅶ. REFERENCES
• Ⅷ. SUMMARY (KOREAN)
• Ⅸ. ACKNOWLEDGMENTS