Guoliang Wang , Lina Xu , Ruize Shi , Yingyue Ye , Baihui Zeng , Xiuqin Yang , Zeyu Liu , Zhen Liu , Shurong Wang , Yuhua Xue , Cheng Li
{"title":"小鼠睑板腺器官型培养模型筛选睑板腺功能障碍相关危险因素","authors":"Guoliang Wang , Lina Xu , Ruize Shi , Yingyue Ye , Baihui Zeng , Xiuqin Yang , Zeyu Liu , Zhen Liu , Shurong Wang , Yuhua Xue , Cheng Li","doi":"10.1016/j.jtos.2023.08.007","DOIUrl":null,"url":null,"abstract":"<div><h3>Purpose</h3><p><span>Meibomian glands (MGs) are crucial for maintaining tear film stability and </span>ocular surface<span> health. Here, we aim to establish a novel organotypic culture model of MGs and explore the risk factors of MG dysfunction (MGD).</span></p></div><div><h3>Methods</h3><p><span><span><span>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, </span>immunofluorescence<span><span>, and qPCR. Lipid accumulation was evaluated by </span>Nile red<span> staining and microscopic examination. Protein expression levels were evaluated by immunofluorescence and </span></span></span>Western blot assay. </span>EdU<span><span> 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 </span>drugs (Dex) into the culture system.</span></p></div><div><h3>Results</h3><p>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<span><span> 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<span>, along with an aberrant increase in Krt10 expression. Further application of this model showed that Dex </span></span>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.</span></p></div><div><h3>Conclusions</h3><p>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.</p></div>","PeriodicalId":54691,"journal":{"name":"Ocular Surface","volume":null,"pages":null},"PeriodicalIF":5.9000,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Organotypic culture model of mouse meibomian gland as a screening platform for risk factors related to meibomian gland dysfunction\",\"authors\":\"Guoliang Wang , Lina Xu , Ruize Shi , Yingyue Ye , Baihui Zeng , Xiuqin Yang , Zeyu Liu , Zhen Liu , Shurong Wang , Yuhua Xue , Cheng Li\",\"doi\":\"10.1016/j.jtos.2023.08.007\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><h3>Purpose</h3><p><span>Meibomian glands (MGs) are crucial for maintaining tear film stability and </span>ocular surface<span> health. Here, we aim to establish a novel organotypic culture model of MGs and explore the risk factors of MG dysfunction (MGD).</span></p></div><div><h3>Methods</h3><p><span><span><span>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, </span>immunofluorescence<span><span>, and qPCR. Lipid accumulation was evaluated by </span>Nile red<span> staining and microscopic examination. Protein expression levels were evaluated by immunofluorescence and </span></span></span>Western blot assay. </span>EdU<span><span> 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 </span>drugs (Dex) into the culture system.</span></p></div><div><h3>Results</h3><p>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<span><span> 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<span>, along with an aberrant increase in Krt10 expression. Further application of this model showed that Dex </span></span>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.</span></p></div><div><h3>Conclusions</h3><p>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.</p></div>\",\"PeriodicalId\":54691,\"journal\":{\"name\":\"Ocular Surface\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":5.9000,\"publicationDate\":\"2023-10-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Ocular Surface\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S1542012423001155\",\"RegionNum\":1,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"OPHTHALMOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Ocular Surface","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1542012423001155","RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"OPHTHALMOLOGY","Score":null,"Total":0}
Organotypic culture model of mouse meibomian gland as a screening platform for risk factors related to meibomian gland dysfunction
Purpose
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.
期刊介绍:
The Ocular Surface, a quarterly, a peer-reviewed journal, is an authoritative resource that integrates and interprets major findings in diverse fields related to the ocular surface, including ophthalmology, optometry, genetics, molecular biology, pharmacology, immunology, infectious disease, and epidemiology. Its critical review articles cover the most current knowledge on medical and surgical management of ocular surface pathology, new understandings of ocular surface physiology, the meaning of recent discoveries on how the ocular surface responds to injury and disease, and updates on drug and device development. The journal also publishes select original research reports and articles describing cutting-edge techniques and technology in the field.
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