Abstract：
OBJECTIVE: Meibomian glands (MGs) are crucial for maintaining tear film stability and ocular surface health. Here, we aim to establish a novel organotypic culture model of MGs and explore the risk factors of MG dysfunction (MGD).
METHODS: We developed a novel organotypic culture model for MGs at the air-liquid interface. The viability and cell proliferation of MGs were assessed using CCK-8, immunofluorescence, and qPCR. Lipid accumulation was evaluated by Nile red staining and microscopic examination. Protein expression levels were evaluated by immunofluorescence and Western blot assay. EdU assay was employed to track the proliferation of acinar cells. The validity of the model was confirmed through culturing MGs from mice of different ages and incorporating certain drugs (Dex) into the culture system.
RESULTS: Utilizing the novel culture model, the MG tissue exhibited sustained viability, cellular division, and continuous production of lipids for a duration of 7 days. Lipid droplets formed were directly visualized using light field microscopy. Through the cultivation of aged mice\'s MGs, it was discovered that aging resulted in diminished proliferation and lipid synthesis, along with an aberrant increase in Krt10 expression. Further application of this model showed that Dex treatment diminished MG\'s proliferation and lipid synthesis. Finally, an in vivo study was conducted to provide additional confirmation of the phenomenon of Dex-induced abnormalities.
CONCLUSIONS: In this study, a stable organotypic culture model of the MGs was established. The organotypic culture model offers a valuable tool to investigate the pathophysiological mechanisms and facilitate drug screening for MG-related diseases.
METHODS: We developed a novel organotypic culture model for MGs at the air-liquid interface. The viability and cell proliferation of MGs were assessed using CCK-8, immunofluorescence, and qPCR. Lipid accumulation was evaluated by Nile red staining and microscopic examination. Protein expression levels were evaluated by immunofluorescence and Western blot assay. EdU assay was employed to track the proliferation of acinar cells. The validity of the model was confirmed through culturing MGs from mice of different ages and incorporating certain drugs (Dex) into the culture system.
RESULTS: Utilizing the novel culture model, the MG tissue exhibited sustained viability, cellular division, and continuous production of lipids for a duration of 7 days. Lipid droplets formed were directly visualized using light field microscopy. Through the cultivation of aged mice\'s MGs, it was discovered that aging resulted in diminished proliferation and lipid synthesis, along with an aberrant increase in Krt10 expression. Further application of this model showed that Dex treatment diminished MG\'s proliferation and lipid synthesis. Finally, an in vivo study was conducted to provide additional confirmation of the phenomenon of Dex-induced abnormalities.
CONCLUSIONS: In this study, a stable organotypic culture model of the MGs was established. The organotypic culture model offers a valuable tool to investigate the pathophysiological mechanisms and facilitate drug screening for MG-related diseases.
摘要：
目的：睑板腺（MGs）对于维持泪膜稳定性和眼表健康至关重要。这里,我们旨在建立一种新型的MGs器官型培养模型，并探讨MG功能障碍（MGD）的危险因素。
方法：我们开发了一种用于气液界面MGs的新型器官型培养模型。使用CCK-8，免疫荧光，和qPCR。通过尼罗红染色和显微镜检查评估脂质积累。通过免疫荧光和Western印迹分析评估蛋白质表达水平。采用EdU测定来追踪腺泡细胞的增殖。通过培养来自不同年龄小鼠的MGs并将某些药物(Dex)掺入培养系统中,证实了模型的有效性。
结果：利用新的文化模型，MG组织表现出持续的活力，细胞分裂,和持续7天的脂质生产。使用光场显微镜直接观察形成的脂滴。通过培养老年小鼠的MGs，发现衰老导致增殖和脂质合成减少，随着Krt10表达的异常增加。该模型的进一步应用表明Dex处理减少了MG的增殖和脂质合成。最后,进行了一项体内研究,以进一步证实Dex诱导的异常现象.
结论：在这项研究中，建立了稳定的MGs器官型培养模型。器官型培养模型为研究MG相关疾病的病理生理机制和促进药物筛选提供了有价值的工具。
方法：我们开发了一种用于气液界面MGs的新型器官型培养模型。使用CCK-8，免疫荧光，和qPCR。通过尼罗红染色和显微镜检查评估脂质积累。通过免疫荧光和Western印迹分析评估蛋白质表达水平。采用EdU测定来追踪腺泡细胞的增殖。通过培养来自不同年龄小鼠的MGs并将某些药物(Dex)掺入培养系统中,证实了模型的有效性。
结果：利用新的文化模型，MG组织表现出持续的活力，细胞分裂,和持续7天的脂质生产。使用光场显微镜直接观察形成的脂滴。通过培养老年小鼠的MGs，发现衰老导致增殖和脂质合成减少，随着Krt10表达的异常增加。该模型的进一步应用表明Dex处理减少了MG的增殖和脂质合成。最后,进行了一项体内研究,以进一步证实Dex诱导的异常现象.
结论：在这项研究中，建立了稳定的MGs器官型培养模型。器官型培养模型为研究MG相关疾病的病理生理机制和促进药物筛选提供了有价值的工具。
