人类结膜外细胞囊泡的丰富协议及其表征
Ismael Romero-Castillo1, Antonio López-García2, Yolanda Diebold2,3
1Ocular Surface Group, Instituto Universitario de Oftalmobiología Aplicada (IOBA), Universidad de Valladolid, Valladolid, Spain. iromeroc@ioba.med.uva.es.
Scientific reports
|November 16, 2024
概括
超过-沉有效地隔离了结膜外细胞囊泡 (EVs). 这些EVs保护角膜细胞免受氧化应激,突出UF沉作为结膜EV丰富的最佳方法.
科学领域:
- 眼科医生 眼科 眼科
- 细胞生物学 细胞生物学
- 生物技术是生物技术.
背景情况:
- 细胞外囊泡 (EVs) 在组织功能中起着至关重要的作用,但它们在角膜保护中至关重要的结膜中的作用尚未得到充分研究.
- 了解结膜EV是推动眼睛表面疾病研究和治疗的关键.
研究的目的:
- 为了比较地从人类结膜表皮和介质细胞中分离和描述EVs.
- 评估不同隔离方法的效率和产量.
- 评估结膜EVs在体外对氧化应激的保护功能.
主要方法:
- 使用差异超离心法 (UC),超过-沉法 (UF-沉) 和超过大小排除色谱法 (UF-SEC) 进行对EV的比较隔离.
- 通过蛋白质含量,大小 (动态光散射),形态 (原子力显微镜) 和标记物表达 (西部斑点) 来描述EVs.
- 在氧化应激条件下对IM-HConEpiC细胞的EV功能影响进行体外试验.
主要成果:
- 这三种方法都成功地回收了EV,而UF沉产生了更高的数量.
- 由UC隔离的EV大于来自UF沉和UF-SEC的EV.
- 结膜性介质 stromal 细胞 (Conj-MSC) 衍生的 EVs 在上皮细胞中显著保护抗氧化应激.
结论:
- 超过-沉是从结膜细胞中分离和丰富细胞外囊泡的最有效方法.
- 结膜EVs,特别是来自介质细胞 stromal 细胞的EVs,具有对氧化应激的保护性质,对角膜健康有意义.
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