PDF(Pubmed)
Abstract：
This study explores the application of the RIP3-caspase3-assay in heterogeneous spheroid cultures to analyze cell death pathways, emphasizing the nuanced roles of apoptosis and necroptosis. By employing directly conjugated monoclonal antibodies, we provide detailed insights into the complex mechanisms of cell death. Our findings demonstrate the assay\'s capability to differentiate between RIP1-independent apoptosis, necroptosis, and RIP1-dependent apoptosis, marking a significant advancement in organoid research. Additionally, we investigate the effects of TNFα on isolated intestinal epithelial cells, revealing a concentration-dependent response and an adaptive or threshold reaction to TNFα-induced stress. The results indicate a preference for RIP1-independent cell death pathways upon TNFα stimulation, with a notable increase in apoptosis and a secondary role of necroptosis. Our research underscores the importance of the RIP3-caspase3-assay in understanding cell death mechanisms in organoid cultures, offering valuable insights for disease modeling and the development of targeted therapies. The assay\'s adaptability and robustness in spheroid cultures enhances its potential as a tool in personalized medicine and translational research.
摘要：
本研究探讨了RIP3-caspase3分析在异质球体培养中的应用，以分析细胞死亡途径。强调细胞凋亡和坏死的细微差别作用。通过使用直接结合的单克隆抗体，我们对细胞死亡的复杂机制提供了详细的见解。我们的发现证明了该测定法区分RIP1非依赖性细胞凋亡的能力，坏死,和RIP1依赖性细胞凋亡，标志着类器官研究的重大进展。此外,我们研究了TNFα对离体肠上皮细胞的影响，揭示了对TNFα诱导的应激的浓度依赖性反应和适应性或阈值反应。结果表明，在TNFα刺激后，对RIP1非依赖性细胞死亡途径的偏好，细胞凋亡明显增加和坏死的次要作用。我们的研究强调了RIP3-caspase3测定在理解类器官培养中细胞死亡机制的重要性。为疾病建模和靶向治疗的发展提供有价值的见解。该测定法在球体培养中的适应性和鲁棒性增强了其作为个性化医学和转化研究工具的潜力。
