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Fig. 5. Kisspeptin-10 inhibits trophoblast outgrowth and migration, but not proliferation in first trimester human villous explant cultures. Villous explants from 6-9 weeks' gestation were maintained in culture for 72 hours in the absence (A,B) or presence of 0.3 µM (C,D) or 1.0 µM (E,F) Kp-10. Identical villi were photographed at 48 or 72 hours. The dark areas are tissue, and sheets of outgrowing cytotrophoblast can readily be observed in the untreated cultures (A,B; arrows mark the limits/boundaries of the outgrowth). Kp-10 treatment profoundly decreases trophoblast outgrowth from the distal end of the villous tips when compared with control villous explants. Magnification: x40. (G-J) Proliferative potential of placental villi was determined by incorporation of BrdU after 24 hours in culture in the absence (G,I) or presence (H,J) of 1 µM Kp-10. Villus sections were stained with an anti-BrdU antibody, which detects cells in the S-phase (G,H), or were labeled with the nuclear stain dapi (I,J). The nonproliferating syncytiotrophoblast (ST) is devoid of anti-BrdU staining (G,H). Cell column (CC) formation by villus explants maintained in the absence or presence of Kp-10 (0.3 µM) was similar. In addition, no significant difference in the proportion of nuclei that incorporated BrdU was detected. Six villi per treatment were examined per placenta and the experiment was repeated three times. Magnification: x400. The extent of migration was increased between 48 and 72 hours under all conditions. Migration distance was reduced already at 0.3 µM Kp-10 at both 48 and 72 hours (K). The higher concentration of 1 µM did not augment the effect. *P<0.05 vs control. VS, villous stroma.
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