在空气-液体界面区分初级人类小气道上皮细胞的协议
Yu Par Aung Myo1, Sarah V Camus2, Margaret A T Freeberg2
1Department of Microbiology and Immunology, Virginia Commonwealth University, Richmond, Virginia, United States.
概括
本研究提供了培养和分化空气管小上皮细胞 (SAECs) 在空气-液体接口 (ALI) 的详细方案. 这种优化的方法可以更好地研究COPD等小型呼吸道疾病.
科学领域:
- 肺研究研究 肺研究
- 细胞培养和分化
- 气道上皮生物学 气道上皮生物学
背景情况:
- 空气液体接口 (ALI) 培养模型模拟了肺部的生理环境.
- 与支气管细胞相比,小气道上皮细胞 (SAEC) 的利用不足.
- SAECs对于研究支气管病理至关重要,并作为膜细胞的替代品.
研究的目的:
- 为ALI提供一个优化,详细的培养和差异化初级人类SAECs的协议.
- 为了解决SAEC ALI文化方法论当前的知识差距.
- 建立一个可靠的模型来研究小呼吸道疾病.
主要方法:
- 优化关键培养条件:通道数,pH,CO2,播种密度和原涂层.
- 开发一种FITC-dextran透性测定,用于评估SAEC屏障完整性.
- 差异化的SAEC暴露于香烟烟雾,LPS,和多I:C) 以验证模型.
主要成果:
- 确定了在ALI的高质量,统一的SAEC差异化关键培养参数.
- 证明了FITC-dextran试验对实验前和后屏障完整性评估的有用性.
- 通过增加细胞因子产生和暴露于刺激时表皮损伤,展示了模型的响应性.
结论:
- 提供的协议成功地弥合了在ALI区分初级人类SAEC的方法差距.
- 这种模型对于推进包括COPD在内的小呼吸道疾病的研究至关重要.
- 优化的SAEC ALI模型有助于研究呼吸道对各种侮辱的反应.
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