Lysophosphatidic Acid Triggers Angiogenesis Through LPA3 Receptor in Human First Trimester Trophoblast

BELTRAME, JS; RIBEIRO MARÍA LAURA; SORDELLI, MS; CAÑUMIL, VA

Orlando - Florida

Congreso; 64th Annual Meeting of the Society for Reproductive Investigation (SRI); 2017

The coordination of vascular processes at the maternal?fetal interface involves extravillous trophoblast differentiation into endovascular trophoblast, facilitating spiral artery remodeling.Lysophosphatidic acid (LPA) is a lipid mediator that regulates female reproductive functions and plays an important role during angiogenesis. Angiogenesis implies the sprouting of existing blood vessels and requires endothelial cells proliferation, migration and tube formation. Using in vitro and in vivo models we investigated the action of LPA in angiogenesis at thematernal?fetal interface. Human first trimester trophoblast cells (HTR-8/SVneo) were seeded on a reduced growth factor membrane matrix (Geltrex®), incubated with LPA (10 -6 M) in th presence of a non-selective LPA3 antagonist (8 Br-LPA 5x10 -6 M), a selective LPA3 antagonist(DGPP 10 -4 M) or a selective LPA1 antagonist (BMT 10 -5 M), and assayed for tube formation during 6h. Tubule length and the number of branches were calculated. MTT Cell Proliferationassay was employed to test trophoblast proliferation ability. Cells were incubated with LPA (10 - 6 M) for 48h. Then, MTT was added for 4h and solubilised formazan is measured spectrophotometrically (540 nm). To test trophoblast migration we employed the wound healing assay. After incubating the cells with LPA (10 -6 M) for 18h we determined the percentage of wound closure. For the in vivo treatment females rats in day 5 of gestation received an intra-uterine dosis of DGPP and the circumferences of blood vessels in theimplantation sites at day 8 of gestation and placentas of day 15 were determined. Results show that LPA stimulates trophoblast capillary response through LPA3 (p