从小鼠新生儿气管呼吸道上皮细胞中优化初级器官样式培养
Yan Han1,2, Rewa Kulkarni2, Stephan Pribitzer3
1Center for Respiratory Biology and Therapeutics, Seattle Children's Research Institute, Seattle, WA, USA.
概括
优化新生儿呼吸道上皮细胞的器官类型培养对于研究肺部发育和损伤至关重要. 与实验室制造的介质相比,商业介质显著改善了细胞增殖,分化,并减少了污染.
科学领域:
- 呼吸系统研究 呼吸系统研究
- 细胞生物学 细胞生物学
- 发育生物学是发展生物学.
背景情况:
- 新生儿呼吸道发育和受伤的了解很少.
- 研究人类病理样本是具有挑战性的.
- 新生儿肺部研究需要有效的ex vivo模型系统.
研究的目的:
- 优化一种用于原始小鼠新生儿气管上皮细胞 (MNTECs) 的器官类型培养的协议.
- 为了比较不同介质条件对MNTEC扩张和分化的有效性.
- 建立一种可靠的技术来研究新生儿呼吸道上皮细胞.
主要方法:
- 鼠新生儿气管上皮细胞 (MNTEC) 的器官类型培养.
- 五种不同的媒体条件的比较 (实验室制造与商业化).
- 状上皮质的量化,基底干细胞 (TP63+),质污染,细胞数量和ALI厚度.
主要成果:
- 与实验室制造的媒体 (20%) 相比,商用媒体的成功率明显更高 (近100%).
- 商业媒体增强了TP63+基底细胞的增殖,并改善了分化成极化,状伪层化气道表皮.
- 商业媒体抑制了受污染的 stromal 细胞的生长.
结论:
- 使用商业媒介进行优化器官类型培养,为研究新生儿呼吸道上皮细胞提供了可靠的方法.
- 这种技术有助于研究新生儿肺部发育,损伤和修复的研究,在野生类型和转基因小鼠中进行.
- 这些发现为我们进一步了解新生儿呼吸道疾病提供了宝贵的工具.
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