Legebeke Jelmer, Horton Katie L, Jackson Claire L, Coles Janice, Harris Amanda, Wai Htoo A, Holloway John W, Wheway Gabrielle, Baralle Diana, Lucas Jane S
School of Human Development and Health, Faculty of Medicine, University of Southampton, Southampton, United Kingdom.
Southampton NIHR Biomedical Research Centre, University Hospital Southampton NHS Foundation Trust, University of Southampton, Southampton, United Kingdom.
Front Cell Dev Biol. 2022 Jun 15;10:907511. doi: 10.3389/fcell.2022.907511. eCollection 2022.
Air-liquid interface (ALI) cell culture of primary airway progenitors enables the differentiation and recapitulation of a pseudostratified epithelium , providing a highly useful tool for researching respiratory health and disease. Previous studies into gene expression in ALI-cultures compared to nasal brushings have been limited in the number of time-points and/or the number of genes studied. In this study physiological and global transcriptomic changes were assessed in an extended 63-day human healthy nasal epithelium ALI-culture period and compared to nasal brushing samples. nasal brushing samples formed distinct transcriptome clusters to ALI-cultured nasal epithelia, with from day 14 onwards ALI samples best matching the samples. Immune response regulation genes were not expressed in the ALI-culture compared to the nasal brushing samples, likely because the cultures lack an airway microbiome, lack airborne particles stimulation, or did not host an immune cell component. This highlights the need for more advanced co-cultures with immune cell representation to better reflect the physiological state. During the first week of ALI-culture genes related to metabolism and proliferation were increased. By the end of week 1 epithelial cell barrier function plateaued and multiciliated cell differentiation started, although widespread ciliation was not complete until day 28. These results highlight that time-points at which ALI-cultures are harvested for research studies needs to be carefully considered to suit the purpose of investigation (transcriptomic and/or functional analysis).
原代气道祖细胞的气液界面（ALI）细胞培养能够实现假复层上皮的分化和重现，为研究呼吸健康与疾病提供了一个非常有用的工具。以往将ALI培养物中的基因表达与鼻刷检样本进行比较的研究，在时间点数量和/或所研究的基因数量方面都受到限制。在本研究中，在长达63天的人类健康鼻上皮ALI培养期内评估了生理和全局转录组变化，并与鼻刷检样本进行了比较。鼻刷检样本形成了与ALI培养的鼻上皮不同的转录组簇，从第14天起，ALI样本与鼻刷检样本最匹配。与鼻刷检样本相比，免疫反应调节基因在ALI培养物中未表达，这可能是因为培养物缺乏气道微生物群、缺乏空气传播颗粒刺激或没有免疫细胞成分。这突出表明需要更先进的包含免疫细胞的共培养，以更好地反映生理状态。在ALI培养的第一周，与代谢和增殖相关的基因增加。到第1周结束时，上皮细胞屏障功能趋于平稳，多纤毛细胞分化开始，尽管直到第28天广泛的纤毛形成才完成。这些结果突出表明，为适合研究目的（转录组学和/或功能分析），需要仔细考虑收获用于研究的ALI培养物的时间点。
