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明胶海绵支持的人鼻黏膜组织培养

Gelatin sponge-supported histoculture of human nasal mucosa.

作者信息

Schierhorn K, Brunnée T, Paus R, Schultz K D, Niehus J, Agha-Mir-Salim P, Kunkel G

机构信息

Department of Clinical Immunology, Free University Berlin, Germany.

出版信息

In Vitro Cell Dev Biol Anim. 1995 Mar;31(3):215-20. doi: 10.1007/BF02639436.

Abstract

Considerable progress has recently been made in the understanding of airway inflammation by cell culture assays and in vivo provocation studies. Inasmuch as ethical considerations limit experimental work in humans, physiologically relevant in vitro models are required to better understand cellular and molecular tissue interactions in human nasal mucosa. Here we describe a human nasal mucosa culture model utilizing a simple gelatin sponge-supported histoculture system at the air-liquid interface. Viable mucosa was preserved for at least 48 h, as shown by morphology and immunohistochemical staining with Ki-67 as marker for proliferation. Pro-inflammatory mediators (kinins, histamine, thromboxane B2, prostaglandin F2 alpha, and substance P) are detectable in serum-containing as well as serum-free culture medium. Incubation with 10(-8) M substance P increases the number of degranulated mast cells after 48 h by 26% (P < 0.01). In this model, biochemical responses can be correlated with histologic alterations of the target tissue. Inflammatory parameters can be examined and compared in various patient groups and different stimulators/inhibitors. This culture method provides a valuable research tool for analyzing all compartments present in nasal mucosa under physiologically relevant conditions, and for studying complex interactions and responses of mucosal cell populations in their natural tissue environment.

摘要

最近,通过细胞培养试验和体内激发研究,在气道炎症的理解方面取得了相当大的进展。由于伦理考虑限制了人体实验工作,因此需要生理相关的体外模型来更好地理解人类鼻黏膜中的细胞和分子组织相互作用。在此,我们描述了一种利用气液界面处简单明胶海绵支撑组织培养系统的人鼻黏膜培养模型。通过形态学以及以增殖标志物Ki-67进行免疫组织化学染色显示,存活的黏膜可保存至少48小时。在含血清以及无血清培养基中均可检测到促炎介质(激肽、组胺、血栓素B2、前列腺素F2α和P物质)。用10(-8) M P物质孵育48小时后,脱颗粒肥大细胞数量增加26%(P < 0.01)。在该模型中,生化反应可与靶组织的组织学改变相关联。可以在不同患者组以及不同刺激物/抑制剂中检查和比较炎症参数。这种培养方法为在生理相关条件下分析鼻黏膜中存在的所有成分,以及研究黏膜细胞群体在其自然组织环境中的复杂相互作用和反应提供了一种有价值的研究工具。

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