Polymorphonuclear leukocyte migration through human amnion membrane.
Polymorphonuclear leukocyte migration through human amnion membrane.
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DOI：
10.1083/jcb.91.2.459
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发表时间：
1981-11
期刊：
影响因子：
--
通讯作者：
Schiffmann E
中科院分区：
文献类型：
--
作者：
Russo RG;Liotta LA;Thorgeirsson U;Brundage R;Schiffmann E
建立了一种新的体外模型来研究人多形核白细胞(PMN)通过活的天然细胞和基质屏障的迁移。人羊膜由一层单层上皮与一层连续的基底膜组成，基底膜与无血管的胶原质相接。将活羊膜放置在塑料室中，膜的两侧各有不同的隔室。中性粒细胞被引入羊膜上皮侧，以及一个米利波尔过滤器(米利波尔公司，马萨诸塞州贝德福德)。被放置在基质一侧。在N-甲酰-甲硫基-亮氨基-苯丙氨酸(FMLP)化学诱导剂的作用下，PMN穿透整个羊膜，收集并计数在过滤器上。羊膜穿透PMN的速率取决于FMLP的浓度(最佳浓度为10(-8)M)以及羊膜上FMLP梯度的斜率。用透射电子显微镜研究了PMN的迁移途径。PMN首先附着于上皮细胞表面，然后在细胞间连接之间渗透。中性粒细胞围绕或通过紧密连接和半桥粒附着体迁移。中性粒细胞穿透基底膜，通过致密的胶原基质迁移。目前的羊膜迁移系统具有体内炎症状态的特征，这是以往任何监测PMN体外迁移的方法所未描述的。以前的方法没有使用天然上皮、整个基底膜或胶原质基质。PMN对这些屏障的穿透发生在正常的炎症反应中，可能涉及到通过人工过滤器的毛孔进行简单迁移所不需要的生化机制。羊膜系统可用于未来对PMN穿透这些屏障的生化和形态学研究，以及随后可能的修复过程。
A new in vitro model has been developed for studying migration of human polymorphonuclear leukocytes (PMN) through living native cellular and matrix barriers. Human amnion membrane consists of a single layer of epithelium bound to a continuous basement membrane interfacing an avascular collagenous stroma. Living amnion was placed in plastic chambers with separate compartments on each side of the membrane. PMN were introduced on the epithelial side of the amnion, and a Millipore filter (Millipore Corp., Bedford, Mass.) was placed against the stromal side. In response to N-formylmethionyl-leucyl- phenylanlanine (FMLP) chemoattractant, PMN penetrated the full thickness of the amnion and were collected and counted on the filter. The rate of PMN traversal of the amnion was dependent on the concentration of FMLP (optimal at 10(-8)M) as well as the slope of the FMLP gradient across the amnion. The route of PMN migration was studied by transmission electron microscopy. PMN first attached to the epithelial surface, then infiltrated between intercellular junctions. PMN migrated around or through tight junction and hemidesmosome attachments. The PMN then penetrated the basement membrane and migrated through the dense collagenous stroma. The present amnion migration system has characteristics of the in vivo inflammatory state not described in any previous method for monitoring PMN migration in vitro. Prior methods have not used native epithelium, whole basement membrane, or collagenous stroma. PMN penetration of these barriers occurs in the normal inflammatory response and probably involves biochemical mechanisms not required for simple migration through the pores of an artificial filter. The amnion system can be useful for future biochemical and morphological studies of PMN penetration of these barriers and possible repair processes that may follow.