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一种对在气液界面(ALI)培养的人原发性呼吸道上皮进行快速评分的方法,用于在研究和个性化医疗环境中评估上皮形态。

A Fast Scoring of Human Primary Respiratory Epithelia Grown at Air-Liquid Interface (ALI) to Assess Epithelial Morphology in Research and Personalized Medicine Settings.

作者信息

Lutsch Christopher T, Feng Longhua, Gómez Hohn Ana, Brandt Lennart, Tamm Stephanie, Janciauskiene Sabina, Stanke Frauke, Jonigk Danny, Dittrich Anna-Maria, Braubach Peter

机构信息

Department of Pediatric Pneumology, Allergology and Neonatology, Hannover Medical School, 30625 Hannover, Germany.

Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH), German Center for Lung Research (DZL), 30625 Hannover, Germany.

出版信息

J Pers Med. 2024 Jan 18;14(1):109. doi: 10.3390/jpm14010109.

Abstract

BACKGROUND

In recent years, increasingly complex ALI protocols involving specialized, albeit laboratory-specific media have been established, while at the same time, many studies compile the data of only a few ALI donors in spite of site-, protocol- and donor-specific differentiation.

METHODS

We describe a simple morphology scoring protocol using histology material derived from epithelia grown on ALI inserts in parallel to other, more complex readouts.

RESULTS

Among more than 100 ALI inserts derived from different donors, significant differences in layer score ( = 0.001) and goblet cell score ( = 0.002) were observed when ALI epithelia derived from explanted lung material were compared to trachea-derived ALI cultures. Cortisol withdrawal for the final 2 days of ALI cultures influenced goblet cell density ( = 0.001).

CONCLUSIONS

While the histology score provides less resolution than FACS- or OMICs- based single cell analyses, the use of a subportion of the ALI epithelia grown on inserts makes it feasible to combine morphology assessment and other readouts of the same insert. This allows us to control for basic ALI morphology in research and personalized medicine settings in order to assess and, if desired, control for the impact of ALI culture protocols, site- and donor-specific influences on outcome of studies of ALI-derived epithelia.

摘要

背景

近年来,已经建立了越来越复杂的ALI方案,这些方案涉及专门的、尽管是特定实验室的培养基,而与此同时,许多研究尽管考虑了部位、方案和供体特异性差异,但仅汇总了少数ALI供体的数据。

方法

我们描述了一种简单的形态学评分方案,该方案使用与其他更复杂读数并行的、源自ALI插入物上生长的上皮组织的组织学材料。

结果

在来自不同供体的100多个ALI插入物中,当将源自移植肺材料的ALI上皮与源自气管的ALI培养物进行比较时,观察到层评分(= 0.001)和杯状细胞评分(= 0.002)存在显著差异。ALI培养最后2天的皮质醇撤离影响了杯状细胞密度(= 0.001)。

结论

虽然组织学评分提供的分辨率低于基于FACS或组学的单细胞分析,但使用在插入物上生长的ALI上皮的一部分,使得将形态学评估与同一插入物的其他读数相结合成为可能。这使我们能够在研究和个性化医学环境中控制基本的ALI形态,以便评估并在需要时控制ALI培养方案、部位和供体特异性影响对ALI衍生上皮研究结果的影响。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/70cd/10817428/6c177b32da2f/jpm-14-00109-g001.jpg

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