METHODS: We established the optimal conditions for coculturing N2a neuronal cells with murine salivary gland organoids (SGOs). After determining the appropriate chemical concentrations, we treated the SGOs cocultured with N2a cells with BoNT type A (BoNT/A). We confirmed the expression of salivary gland-related genes and proteins using real-time polymerase chain reaction (PCR) and immunofluorescence staining.
RESULTS: The SGOs cocultured with N2a cells showed that the dendrites or axons of neuronal cells were in contact with the outermost layer of the SGOs. When we applied acetylcholine and neostigmine to the coculture systems, the mRNA expression of Aqp5 and Bhlha15, associated with salivary gland secretory cells, increased. However, this effect was reversed when BoNT/A was applied, as confirmed through real-time PCR.
CONCLUSIONS: We found that the coculture system of SGOs and N2a neuronal cells can potentially serve as a potency testing platform for BoNT.
METHODS: NA Laryngoscope, 134:2697-2704, 2024.
方法:我们建立了将N2a神经元细胞与鼠唾液腺类器官(SGO)共培养的最佳条件。在确定合适的化学浓度后,我们用BoNTA型(BoNT/A)处理与N2a细胞共培养的SGO。我们使用实时聚合酶链反应(PCR)和免疫荧光染色证实了唾液腺相关基因和蛋白质的表达。
结果:与N2a细胞共培养的SGO表明,神经元细胞的树突或轴突与SGO的最外层接触。当我们将乙酰胆碱和新斯的明应用于共培养系统时,与唾液腺分泌细胞相关的Aqp5和Bhlha15的mRNA表达,增加。然而,当应用BoNT/A时,这种效果被逆转,通过实时PCR证实。
结论:我们发现SGO和N2a神经元细胞的共培养系统可以潜在地充当BoNT的效力测试平台。
方法:NA喉镜,2024.