Zaga-Clavellina CV, López-Vancell R, Maida-Claros R, Beltrán-Montoya J, Vadillo-Ortega F

Language: Spanish
References: 22
Page: 162-171
PDF size: 312.37 Kb.

ABSTRACT

Objective: This study was designed to validate and characterize a culture model of human choriamniotic membranes (HCM) that keeps their viability, integrity and capacity to reproduce a response to several stimulus associated to an infectious process, as the tissue that separates the fetal and maternal compartment.

Material and methods: We use HCM obtained after delivery by elective cesarean section. Women with 37-40 weeks of gestation without evidence of active labor or presence of clinical an microbiological signs of intrauterine/vaginal infection. The membranes were mounted in transwell devices, allowing testing two independent compartments (chorion and amnion) by physically separating the upper and lower chambers. 500 ng/mL of lipopolysaccharide was added to amniotic or chorionic surface and secretions of TNFα was measured in both compartments by specific enzyme-linked immunosorbent assays and Metalloproteinase-9 (MMP-9) secretions after stimulation with 5 ng/mL of IL-1β.
Results: The viability test showed that the different cellular populations of the HCM keep their metabolic viability along 96 h of culture. The integrity parameters showed that the stay without morphologic and structural changes. Functional markers showed that membranes responded differentially to IL-1β stimulus; production of MMP-9 in chorion reached its maximum value at 4 h, while amnion reached it at 24 h. The selective stimulation of chorioamnion with lipopolysaccharide induced a differential synthesis of TNFα; the chorion was the principal producer with approximately 60% of total TNFα.
Conclusions: The experimental model allows to study qualitatively and quantitatively the contribution of different cellular regions of the HCM, and its response to differential stimulation with immunologic agents.

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