Isolation of canine trophoblasts and chemokine response to stimulation with TLR agonists and B. canis antigens.

(A) Trophoblasts isolated from term placentas of healthy dogs stained positive for cytokeratin 7 and negative for vimentin. (B) Chemokine response of canine trophoblasts isolated from 6 term canine placentas stimulated for 24 h with 5 μg/ml Pam3CSK4 (Pam), 5 μg/ml of E. coli lipopolysaccharide (LPS), 1 μg/ml of S. enterica flagellin (Flag) and 10⁹ colony forming units (CFU) /mL of heat-killed Brucella canis (HKBc). Values in B are means ± SD of duplicate measurements from three independent experiments. Asterisks indicate significant differences as compared to unstimulated (control) cells (* p<0.05; *** p<0.001, ANOVA followed by Dunnett᾽s test).