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. Author manuscript; available in PMC: 2020 Aug 1.
Published in final edited form as: Reprod Toxicol. 2019 May 30;87:100–107. doi: 10.1016/j.reprotox.2019.05.064

Figure 5. THC, CBD and CBN compromise trophoblast invasion and MMP gene expression.

Figure 5.

A. A schematic illustration of the Boyden chamber assay that was used to evaluate the effects of THC, CBD or CBN on trophoblast-THESC interaction. First, THESCs were plated and decidualized for 7 days with or without 0.5 μM THC, CBD or CBN. Then, martigel-coated transwells layered with HTR8 cells were placed to the chamber as shown. Trophoblast invasion was assessed 12 hours later following staining with crystal voilet. B. Undecidualized TESECs induced lesser HTR8 cell migration (as indicated in violet color) (image a) compared to decidualized TESECs (MEF alone) (image b). The presence of THC (image c), CBD (image d) or CBN (image e) during decidualization reduced decidualization-induced HTR8 cell migration. The scale bars in images represent 600 μm. C. Invading HTR8 cells were counted after staining with crystal violet. The increase in invading HTR8 cells associated with THESC decidualization was significantly reduced by the presence of THC, CBD or CBN. D. HTR8 cells were cultured in conditioned media collected from undecidualized control and decidualized THESCs with or without THC, CBD or CBN treatment. The significant increase in mRNA levels of MMP2 and MMP9 in HTR8 cells associated with THSEC decidualization was significantly reduced by the presence of THC, CBD or CBN in conditioned media, suggesting that decidualization creates a microenvironment conducive for TB invasion and that CBs can compromise the formation of this microenvironment. *indicates significant increase (p ≤ 0.05) in invading HTR8 cells in the presence of decidualized THSECs compared to undecidualized THSECs or significant increase (p ≤ 0.05) in mRNA levels of MMP2 or MMP9 in HTR8 cells cultured in conditioned media from decidualized THSECs compared to undecidualized THSECs. $indicates significant decrease (p ≤ 0.05) in invading HTR8 cells in the presence of decidualized THSECs treated with THC, CBD or CBN versus no CB treatment or significant decrease (p ≤ 0.05) in mRNA levels of MMP2 or MMP9 in HTR8 cells cultured in conditioned media from decidualized THESCs treated with THC, CBD or CBN versus no CB treatment. Data shown are means ± SEM of four independent experiments performed in duplicate in each experiment. N and D in panels C and D represent undecidualized and decidualized THESCs, respectively.