Effects of glucose and fatty acids interactions on glucose metabolism pathways in human trophoblasts [article]

Carina Bertoldi Franco, University, The Australian National, University, The Australian National
2015
Obesity, with its associated co-morbidities of diabetes, hyperlipidaemia and low-grade inflammation, has high prevalence in women of child-bearing age. Of further concern, it is associated with increased risk of adverse pregnancy outcomes. Considering the key role of the placenta in fetal development, alterations to its structure and function in response to the adverse metabolic milieu of maternal obesity must be important. The trophoblast is a key cell within the placental maternal-fetal
more » ... r. The research aims were to evaluate the effects of elevated glucose supply, in the absence and presence of non-esterified fatty acids (NEFA), on pathways of glucose metabolism and trophoblast morphology in human trophoblasts in primary culture. Methods: Trophoblasts were established in primary culture from human term placentas of normal pregnancies. The isolation protocol was optimised to improve trophoblast yield and purity. Experiments were performed at both 36 h (short-term) and 5 days (long-term) after trophoblast isolation, when most cells were, respectively, cytotrophoblasts and syncytiotropblasts. Syncytialisation was promoted by adding epidermal growth factor (EGF; 10 ng/ml) to the media. Metabolism experiments were performed over 2 h at diverse glucose concentrations in the absence or presence of NEFA (0.25 mM, palmitate: oleate ratio; 1:1). Pathways of glucose uptake, glycolysis, glucose oxidation, total and partial palmitate oxidation and glycogen synthesis were assessed using radio-labelled tracers. Lactate production and glycogen content were measured enzymatically. The effects of EGF and NEFA on trophoblast morphology, lipid droplet formation and triacylglycerol (TG) content were also assessed. Results: The improved protocol increased the mean (± SD) trophoblast yield to 266 ± 141 x 10⁶ cells, and purity to 98%. In short-term cultures, rates of trophoblast glucose uptake and glycogen synthesis increased linearly with increasing glucose concentrations; whereas, rates of glycolysis, lactate rele [...]
doi:10.25911/5d6e4af2a46b5 fatcat:lk6tluj4czgnzlxca43lskgjte