Lipopolysaccharide from e coli o111 b4

To as sess CHDP ex pres sion in re sponse to in fec tion, an in vitro ovine pla cen tal model was em ployed. AH1 tro phoblast cells were grown to 80% con flu ency overnight in a 24 well plate, and sub se quently ex posed to LPS or in fected with live W. chon drophila. For LPS ex po sures, AH1 cells were treated in du pli cate with LPS (500 ng/ mL, lipopolysac cha ride from E.coli O111:B4, Sigma, L2630-10 mg) which was dis solved in ul tra pure wa ter and stored at -80 °C (n = 4). AH1 cells were also in fected with W. chon drophila at a mul ti plic ity of in fec tion (MOI) of 0.1, 1 and 10 (equiv a lent of 1 in clu sion form ing unit (IFU) per 10 cells, 1 IFU per cell and 10 IFU per cell, re spec tively) or ex posed to UV ir ra di ated or gan isms at MOI 10 (n = 3). Cells were main tained in a heated, hu mid i fied in cu ba tor at 37 °C with 5% CO for 48 h dur ing treat ment. For Vi t a min D ex po sures, 1α,25di hy drox yvi t a min D (cal citriol, Enzo Life Sci ences, Ex eter, United King dom) was dis solved in ab solute al co hol at a stock con cen tra tion of 1 mM and stored at -80 °C. AH1 cells were ex posed to Vi t a min D in du pli cate at con cen tra tions of 1 nM, 10 nM and 100 nM for 24 h and in all treat ments, a ve hi cle con trol was per formed (1 μl/ mL EtOH) (n = 3).