Assessment of Placental Sodium-Independent Leucine Uptake and Transfer in Trophoblast Cells.

Abstract

The placenta maintains the balance between nutrition and growth control of the fetus through selective and regulated supply of macronutrients such as carbohydrates, proteins, lipids, and critical micronutrients. Perturbations in the balanced supply of nutrients as found in gestational diseases and altered fetal development have been associated with changes in amino acid transport proteins, such as the System L amino acid heterodimeric exchangers LAT1/SLC7A5 and LAT2/SLC7A8. Syncytiotrophoblasts (STB) form the crucial cell layer at the placental barrier coordinating the transfer of essential amino acids such as leucine from the maternal to the fetal circulation. The System L-mediated leucine transport across the placental barrier is a Na+-independent process against a counter-directed gradient, maintained by a tightly regulated interplay between accumulative transporters, exchangers, and facilitators.The two methods described here allow to standardize and characterize the uptake kinetics of leucine in conventionally cultured BeWo cells and the transfer of leucine across the placental cell barrier using a BeWo monolayer in the Transwell® system.