Page 21 - HKSEMR2020 Programme book

Poster Presentation (Basic Science) Abstracts

Identification of pharmacologically active molecules that suppress

spheroid attachment and embryo implantation in vitro and in vivo

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Xian Chen , Raymond HW Li , Ernest HY Ng , William SB Yeung , Kai-Fai Lee
1. Department of Obstetrics and Gynaecology, LKS Faculty of Medicine, The University of Hong Kong, Hong Kong
SAR, China; 2. Shenzhen Key Laboratory of Fertility Regulation. The University of Hong Kong-Shenzhen Hospital,
Haiyuan 1st Road, Futian District, Shenzhen, 518053, China

Introduction / Background / Objectives: pregnant ICR female mice were trans-cervical transferred with
3 μL chemical solution (positive control: Misoprostol, 250 μg/kg;
Human endometrium is receptive to embryo implantation during
the mid-secretory phase of the menstrual cycle. In vitro spheroid- vehicle controls: 5% DMSO and PBS; selected LOPAC molecules)
endometrial epithelial cells co-culture model has long been used on 2.5 dpc and the implantation sites on 5.5 dpc were evaluated.
to study the attachment of spheroids (blastocyst surrogates) onto
endometrial epithelial cells. We hypothesized that the molecules Results / Outcomes:
from the library of 1280 pharmaceutically active compounds
(LOPAC) can effectively modulate embryo implantation and Screening of 1,280 LOPAC identified 173 molecules (p < 0.05)
potentially be used for emergency contraception. significantly suppressed the spheroid attachment rate. Eight
potential molecules were identified with low LC 50 and three of
them were found to suppress spheroid attachment in RL95-2
Methods: cells without significantly inhibiting the cell viability. Two out of
three, Molecule 14 (100 μg/kg) and Molecule 46 (15 and 150 μg/
We established a high-throughput in-vitro spheroid-endometrial
epithelial (Ishikawa) cells coculture model in a 96-well plate kg), were found to significantly decrease (p < 0.05) the number
format. Endometrial cells were grouped as negative control, vehicle of implantation sites on the treated sides (5.8±2.4 for Molecule
control, positive control (PRI-724, 10 μM, 24h) or treatment groups 14, 5.0±2.3 and 5.8±1.9 for Molecule 46, respectively) when
(LOPAC, 10 μM, 24h). The trophoblastic BeWo spheroids were compared with non-treated sides (8.0±1.5 for Molecule 14,
generated by AggreWellTM and labeled with a fluorescent dye for 8.3±1.7 and 8.8±2.3 for Molecule 46, respectively) (n = 11-12).
quantitation of the number of the seeded and attached spheroids
by a fluorescent plate reader. The loosely bound spheroids were Conclusion:
washed off with PBS. The toxicity of molecules was detected by
XTT cell viability assay. Molecules with low toxicity that suppress Molecule 14 and 46 can suppress spheroid attachment and
spheroid attachment were studied at lower concentrations in embryo implantation in vitro and in vivo.
another receptive endometrial epithelial cell line (RL95-2). The

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