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Article

Effects of Natural Progesterone and Synthetic Progestin on Germ Layer Gene Expression in a Human Embryoid Body Model

1
Department of Obstetrics and Gynecology, Seoul National University College of Medicine, Seoul 03080, Korea
2
Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University College of Medicine, Seoul 03080, Korea
3
Center for Reproductive Medicine and Infertility, Weill Medical College of Cornell University, New York, NY 10065, USA
*
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Int. J. Mol. Sci. 2020, 21(3), 769; https://doi.org/10.3390/ijms21030769
Received: 15 November 2019 / Revised: 16 January 2020 / Accepted: 17 January 2020 / Published: 24 January 2020
Natural progesterone and synthetic progestin are widely used for the treatment of threatened abortion or in in vitro fertilization (IVF) cycles. This in vitro study aimed to assess whether the treatment with natural progesterone or synthetic progestin influences the germ layer gene expression on the early human embryonic development using human embryonic stem cells (hESCs)-derived embryoid bodies (hEBs) as a surrogate of early stage human embryonic development. Human EBs derived from hESCs were cultured for nine days, and were treated with natural progesterone (P4) or synthetic progestin, medroxyprogesterone acetate (MPA) at 10–7 M for five days. To reverse the effects of treatment, mifepristone (RU486) as progesterone antagonist was added to the hEBs for four days starting one day after the initiation of treatment. Mouse blastocysts (mBLs) were cultured in vitro for 24 h, and P4 or MPA at 10−7 M was treated for an additional 24 h. The treated embryos were further transferred onto in vitro cultured endometrial cells to evaluate chorionic gonadotropin (CG) expression. To analyze the effects of P4 or MPA, the expression of differentiation genes representing the three germ layers was investigated, GATA-binding factor 4 (GATA4), α-fetoprotein (AFP), hepatocyte nuclear factor (HNF)-3β, hepatocyte nuclear factor (HNF)-4α (endoderm), Brachyury, cardiac actin (cACT) (mesoderm), and Nestin (ectoderm), using quantitative reverse transcription PCR (qRT-PCR) and immunostaining. Significantly lower expressions of HNF-3β, HNF-4α, Brachyury, and Nestin were observed in MPA-treated hEBs (all p < 0.05), which was negated by RU486 treatment. This inhibitory effect of MPA was also observed in mouse embryos. Conclusively, the effects of natural progesterone and synthetic progestin may differ in the germ layer gene expression in the hEB model, which suggests that caution is necessary in the use of progestogen. View Full-Text
Keywords: progesterone; progestin; human embryoid body; mouse embryo; early development progesterone; progestin; human embryoid body; mouse embryo; early development
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MDPI and ACS Style

Kim, Y.Y.; Kim, H.; Suh, C.S.; Liu, H.-C.; Rosenwaks, Z.; Ku, S.-Y. Effects of Natural Progesterone and Synthetic Progestin on Germ Layer Gene Expression in a Human Embryoid Body Model. Int. J. Mol. Sci. 2020, 21, 769. https://doi.org/10.3390/ijms21030769

AMA Style

Kim YY, Kim H, Suh CS, Liu H-C, Rosenwaks Z, Ku S-Y. Effects of Natural Progesterone and Synthetic Progestin on Germ Layer Gene Expression in a Human Embryoid Body Model. International Journal of Molecular Sciences. 2020; 21(3):769. https://doi.org/10.3390/ijms21030769

Chicago/Turabian Style

Kim, Yoon Y., Hoon Kim, Chang S. Suh, Hung-Ching Liu, Zev Rosenwaks, and Seung-Yup Ku. 2020. "Effects of Natural Progesterone and Synthetic Progestin on Germ Layer Gene Expression in a Human Embryoid Body Model" International Journal of Molecular Sciences 21, no. 3: 769. https://doi.org/10.3390/ijms21030769

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