Establishment and comparison of air-liquid interface culture systems for primary and immortalized swine tracheal epithelial cells.
Establishment and comparison of air-liquid interface culture systems for primary and immortalized swine tracheal epithelial cells.
复制标题
原代与永生化猪气管上皮细胞气液界面培养体系的建立与比较
DOI：
10.1186/s12860-018-0162-3
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发表时间：
2018-06-28
期刊：
影响因子：
--
通讯作者：
Feng Z
中科院分区：
文献类型：
--
作者：
Wang H;He L;Liu B;Feng Y;Zhou H;Zhang Z;Wu Y;Wang J;Gan Y;Yuan T;Wu M;Xie X;Feng Z
背景空气-液体界面(ALI)系统允许建立比其他培养系统更能代表体内的培养环境。它们有助于进行呼吸道上皮细胞作为药物渗透屏障的机制研究，并可用于研究宿主和呼吸道病原体之间的相互作用。结果建立了原代和永生化猪气管上皮细胞ALI培养体系，并首次系统比较了这两种体系的分化能力和免疫功能。在ALI培养条件下，永生化的STEC和原代STEC可存活至少40d，并形成紧密连接并分化为复层细胞。它们都具有产生粘蛋白和炎性细胞因子以及发育纤毛的完全能力。然而，与具有异质性形态的原代STEC相反，ALI培养的永生化STEC系似乎是同源的群体。ALI培养的原代STEC形成紧密连接的能力明显优于永生化STEC。此外，ALI培养的永生化STEC纤毛明显高于原代STEC，但表达持续时间短于原代STEC。结论本研究首次建立了具有完整纤毛分化和炎性细胞因子产生能力的ALI原代STEC和永生化STEC细胞系，并观察到两者在形态、紧密连接和纤毛形成等方面的差异。这两个系统将成为药物筛选研究的重要工具，以及详细分析宿主和呼吸道病原体之间的相互作用。
BackgroundAir-liquid interface (Ali) systems allow the establishment of a culture environment more representative of that in vivo than other culture systems. They are useful for performing mechanistic studies of respiratory epithelial cells as drug permeation barriers and can be used to study the interactions between hosts and respiratory pathogens. However, there have been few studies concerning Ali cultures of primary swine tracheal epithelial cells (STECs) and an immortalized STEC line, and the differences between these two systems remain poorly defined.ResultsIn this study, we established Ali culture systems for primary STECs and for immortalized STEC line, and we systematically compared the differentiation capacities and immunological functions of these systems for the first time. Under Ali culture conditions, immortalized STEC line and primary STECs could survive for at least forty days, formed tight junctions and differentiated into stratified cells. They both possessed complete abilities to produce mucin and inflammatory cytokines and develop cilia. However, in contrast to primary STECs, which had a heterogeneous morphology, Ali-cultured immortalized STEC line appeared to be a homogenous population. The formation of tight junctions in Ali-cultured primary STECs was superior to that in immortalized STEC line. In addition, cilia in Ali-cultured immortalized STEC line were more pronounced, but their duration of expression was shorter than in primary STECs.ConclusionsAli-cultured primary STECs and immortalized STEC line systems possessing complete abilities to undergo ciliary differentiation and inflammatory cytokine production were established for the first time in this study, and several differences in morphology and the formation of tight junctions and cilia were observed between these two systems. These two systems will be important tools for drug screening studies, as well as for detailed analyses of the interactions between hosts and respiratory pathogens.
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DOI：
10.1056/nejmra0910061
发表时间：
2010-12-02
期刊：
The New England journal of medicine
影响因子：
--
作者：
Fahy JV;Dickey BF
通讯作者：
Dickey BF
影响因子：
2.6
作者：
Eisele NA;Anderson DM
通讯作者：
Anderson DM
影响因子：
2.7
作者：
Hwang, Mi-Hyun;Chang, Zhi-Qiang;Park, Seung-Chun
通讯作者：
Park, Seung-Chun
影响因子：
3.8
作者：
Krunkosky, Thomas M.;Jordan, Jarrat L.;Krause, Duncan C.
通讯作者：
Krause, Duncan C.
DOI：
10.1016/j.ejpb.2005.02.010
发表时间：
2005-07-01
影响因子：
4.9
作者：
Forbes, B;Ehrhardt, C
通讯作者：
Ehrhardt, C