Effect of Obesity on Mitochondrial Function and Hypoxia Susceptibility in Human Syncytiotrophoblasts

Placentae were collected from women (n=28) with a range of BMI (18-45) after C-section at term with no labor. Villous cytotrophoblasts were isolated from placentae by tryspsin/DNAse digestion and purified by separation on a Percoll gradient and cultured for 72 hrs to form syncytiotrophoblasts (1). Cellular bioenergetics was measured using a Seahorse Bioscience XF24 Analyzed as described (2). Oxygen consumption rate (OCR) and extracellular acidification rate (ECAR), a measure of glycolysis, were measured (Figure 2). In addition, basal and oligomycin treated (ATP synthase inhibited) ECAR were determined to assay resting glycolysis and maximum glycolytic rates. To simulate the hypoxic environment known to be involved in the pathogenesis of obesity, we treated trophoblasts from the 48 to 72hr interval with 50 μM desferroxamine (DFO), a hypoxia mimetic. Trophoblast OCR was measured using the XF24.