Naaz Rimshia, Kuruburu Mahadevaswamy G, Leihang Zonunsiami, Bovilla Venugopal R, Rajashetty Rajalakshmi, Madhusetty Ramya C, Vaagesh Vijaya Y, Madhunapantula SubbaRao V
Center of Excellence in Molecular Biology and Regenerative Medicine, Department of Biochemistry, JSS Medical College, Faculty of Medicine, JSS Academy of Higher Education and Research, Mysuru, Karnataka 570015, India.
Department of Physiology, JSS Medical College, Faculty of Medicine, JSS Academy of Higher Education and Research, Mysuru, Karnataka 570015, India.
J Biol Methods. 2025 Aug 28;12(3):e99010071. doi: 10.14440/jbm2025.0016. eCollection 2025.
cell culture is essential for elucidating various signaling mechanisms and screening pharmacological agents to assess their safety and efficacy. However, cell proliferation and survival in culture can be significantly influenced by variations in the composition of the culture medium. For instance, variations in glucose and fetal bovine serum (FBS) concentrations can impact cell viability. Despite this, only a few studies have examined the impact of varied FBS and glucose concentrations in culture media on cell viability.
This study investigated the mechanisms and cellular effects of glucose and FBS deprivation in glioblastoma cell lines.
We systematically evaluated the impact of FBS and glucose deprivation on the proliferation and survival of rat C6 and human U-87 MG glioblastoma cell lines.
Glucose deprivation (0 mg/dL) significantly reduced the viability of C6 cells and moderately lowered the viability of U-87 MG cells, with partial recovery upon glucose supplementation (100 mg/dL, 400 mg/dL). Notably, FBS deprivation (0%) exerted a more profound effect, inducing the accumulation of reactive oxygen species and extensive cell death in both cell lines. Restoration of FBS (1, 2, 4, 6, 8, and 10%) recovered cell viability and reduced oxidative stress. Furthermore, both glucose and FBS deprivation altered antioxidant enzyme expression and mitochondrial function. Glucose and FBS deprivation also differentially affected protein kinase B phosphorylation, suggesting metabolic stress-induced signaling modulation.
These findings highlight the differential responses of glioblastoma cells to glucose and FBS deprivation and underscore the importance of standardizing culture conditions, especially serum and glucose levels, when designing experiments involving glioblastoma cells.
细胞培养对于阐明各种信号传导机制以及筛选药物制剂以评估其安全性和有效性至关重要。然而，培养基成分的变化会显著影响培养中的细胞增殖和存活。例如，葡萄糖和胎牛血清（FBS）浓度的变化会影响细胞活力。尽管如此，只有少数研究考察了培养基中不同FBS和葡萄糖浓度对细胞活力的影响。
本研究探讨了胶质母细胞瘤细胞系中葡萄糖和FBS剥夺的机制及细胞效应。
我们系统评估了FBS和葡萄糖剥夺对大鼠C6和人U-87 MG胶质母细胞瘤细胞系增殖和存活的影响。
葡萄糖剥夺（0 mg/dL）显著降低了C6细胞的活力，并适度降低了U-87 MG细胞的活力，补充葡萄糖（100 mg/dL，400 mg/dL）后部分恢复。值得注意的是，FBS剥夺（0%）产生了更深远的影响，在两种细胞系中均诱导活性氧的积累和广泛的细胞死亡。恢复FBS（1%、2%、4%、6%、8%和10%）可恢复细胞活力并降低氧化应激。此外，葡萄糖和FBS剥夺均改变了抗氧化酶表达和线粒体功能。葡萄糖和FBS剥夺还对蛋白激酶B磷酸化产生不同影响，提示代谢应激诱导的信号调节。
这些发现突出了胶质母细胞瘤细胞对葡萄糖和FBS剥夺的不同反应，并强调了在设计涉及胶质母细胞瘤细胞的实验时标准化培养条件的重要性，尤其是血清和葡萄糖水平。
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