Department of Immunobiochemistry, Instituto Nacional de Perinatología Isidro Espinosa de los Reyes, Mexico City, Mexico.
Department of Infectology and Immunology, Instituto Nacional de Perinatología Isidro Espinosa de los Reyes, Mexico City, Mexico.
Methods Mol Biol. 2024;2781:27-37. doi: 10.1007/978-1-0716-3746-3_3.
Leukocyte infiltration into the maternal-fetal interface is a consequence of the robust inflammation in the gestational tissues during term labor and preterm labor with or without infection. During pregnancy, the fetal membranes act as a physical barrier that isolates the fetus into the amniotic cavity, keeping it in an optimal environment for its development. In addition, the fetal membranes possess immunological competencies such as the secretion of cytokines and chemokines in response to different stimuli. Clinical and experimental evidence indicates that these tissues are involved in the extensive chemotaxis of immune cells in normal or pathological conditions.Few studies have evaluated the chemotactic capacities of the fetal membranes considering that this tissue is composed of two adjacent tissues, the amnion and the chorion, which have different characteristics. Although these tissues function as a unit, their response is complex since there is an interaction between them, where each tissue contributes differently. The protocol described here allows us to evaluate the in vitro chemotactic capacities of fetal membranes in response to various applied stimuli, considering the contribution of each of their components (amnion and choriodecidua) using a Boyden chamber assay and phenotyping the chemo-attracted leukocytes by flow cytometry.
白细胞浸润到母胎界面是足月分娩和有或没有感染的早产时妊娠组织中强烈炎症的结果。在怀孕期间,胎膜作为一种物理屏障,将胎儿隔离在羊膜腔中,为其发育提供最佳环境。此外,胎膜具有免疫能力,例如对不同刺激物分泌细胞因子和趋化因子。临床和实验证据表明,这些组织参与正常或病理条件下免疫细胞的广泛趋化作用。很少有研究评估胎膜的趋化能力,因为该组织由两个相邻的组织组成,即羊膜和绒毛膜,它们具有不同的特征。尽管这些组织作为一个整体发挥作用,但由于它们之间存在相互作用,每个组织的贡献不同,因此它们的反应非常复杂。这里描述的方案允许我们使用 Boyden 室测定法评估对各种应用刺激物的胎膜的体外趋化能力,并考虑到它们各自组成部分(羊膜和绒毛膜蜕膜)的贡献,通过流式细胞术对趋化性白细胞进行表型分析。