OS041. XXXX protects normal integration of the trophoblast into endothelial cellular networks in an in vitro model of @entity1468 .

@entity1 @entity2299 @entity310 @entity1674 @entity409 @entity1468 @entity1675
INTRODUCTION: Failure of the trophoblast to appropriately invade uterine spiral arteries is thought to be an initiating event in @entity1468 , a disorder associated with @entity409 . @entity2299 characterised by low plasma levels of high density lipoproteins (HDL) and elevated @entity1674 has also been described in @entity1468 . The pro-inflammatory cytokine @entity310 inhibits trophoblast invasion of uterine endothelial cells. Previous work using an in vitro JEG-3 cell/Uterine endothelial cell co-culture model investigated the effect of apoliopoprotein A-I, the main apolipoprotein component of HDL, on trophoblast incorporation into endothelial tubules in the presence and absence of @entity310 . These effects are now investigated using the @entity1 invasive trophoblast cell line HTR-8/SVneo. OBJECTIVES: This study asks if @entity1675 , which has established anti-inflammatory properties, can protect against the deleterious effect of @entity310 on trophoblast-endothelial cell interactions. METHODS: The in vitro trophoblast-uterine endothelial cell co-culture model was used to investigate the effect of @entity1675 on trophoblast incorporation into endothelial tubules in the presence and absence of @entity310 . Uterine endothelial cells were pre-incubated with lipid free @entity1675 (final @entity1675 concentration 1 mg/mL) for 16h prior to seeding on matrigel coated plates. Tubules formed within 4h. Fluorescence-labelled HTR-8/SVneo trophoblast cells were then co-cultured with the endothelial cells  @entity310 (final concentration of 0.2ng/mL). Bright field and fluorescent images were captured after 24h. The effect of @entity310 on HTR-8/SVneo cell invasion was quantified with Image J software. Integration of HTR-8/SVneo trophoblast cells into uterine endothelial tubular networks was also imaged using live cell imaging techniques (Zeiss Axiovert). RESULTS: @entity310 inhibited HTR-8/SVneo (trophoblast) cell integration into endothelial tubular structures by 24.1  3.7% p<0.001. This effect was reversed when the endothelial cells were pre-incubated for 16h with lipid free @entity1675 (p<0.001 compared to non-incubated cells). Live cell images of the co-culture clearly demonstrate a disruption to the normal integration of trophoblast into endothelial tubular structures in the presence of @entity310 . The protective effect conferred by pre-incubation of endothelial cells with @entity1675 against @entity310 is also clearly visible. CONCLUSION: @entity1675 has been shown to enhance trophoblast-endothelial cell integration in the presence of a pro-inflammatory stimulus. A healthy lipid profile may affect pregnancy outcomes by priming endothelial cells in preparation for trophoblast invasion.
