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Updated: Jul 17, 2025
Isolation of Human Endometrial Stromal Cells for In Vitro Decidualization
Published on: September 1, 2018
miRNAs associated with endoplasmic reticulum stress and unfolded protein response during decidualization
Elizabeth Soczewski1, José Martin Murrieta-Coxca2, Lucas Miranda1
1CONICET, Universidad de Buenos Aires, Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales IQUIBICEN, Buenos Aires, Argentina.
Research Question:
Do microRNAs (miRNAs) play a role in regulating endoplasmic reticulum stress (ERS) and unfolded protein response (UPR) in decidualized cells and endometrium associated with reproductive failures?
Design:
Endometrial stromal cell line St-T1b was decidualized in vitro with 8-Br-cAMP over 5 days, or treated with the ERS inducer thapsigargin. Expression of ERS sensors, UPR markers and potential miRNA regulators was analysed by quantitative PCR. Endometrial biopsies from patients with recurrent pregnancy loss (RPL) and recurrent implantation failure (RIF) were investigated for the location of miRNA expression.
Results:
Decidualization of St-T1b cells resulted in increased expression of ERS sensors including ATF6α, PERK and IRE1α, and the UPR marker, CHOP. TXNIP, which serves as a link between the ERS pathway and inflammation, as well as inflammasome NLRP3 and interleukin 1β expression increased in decidualized cells. An in-silico analysis identified miR-17-5p, miR-21-5p and miR-193b-3p as miRNAs potentially involved in regulation of the ERS/UPR pathways and inflammation associated with embryo implantation. Their expression decreased significantly (P ≤ 0.0391) in non-decidualized cells in the presence of thapsigargin. Finally, expression of the selected miRNAs was localized by in-situ hybridization in stromal and glandular epithelial cells in endometrial samples from patients with RPL and RIF. Expression in stroma cells from patients with RPL was lower in comparison with stroma cells from patients with RIF.
Conclusions:
Decidualization in St-T1b cells is accompanied by ERS/UPR processes, associated with an inflammatory response that is potentially influenced by miR-17-5p, miR-21-5p and miR-193b-3p. These miRNAs are expressed differentially in stromal cells from patients with RPL and RIF, indicating an alteration in regulation of the ERS/UPR pathways.
Insights
MicroRNAs (miRNAs) like miR-17-5p, miR-21-5p, and miR-193b-3p may regulate endoplasmic reticulum stress (ERS) and inflammation in decidualized cells. Their altered expression in patients with reproductive failure suggests a role in implantation issues.
Area of Science:
- Reproductive biology
- Molecular biology
- Cellular stress response
Background:
- Endoplasmic reticulum stress (ERS) and unfolded protein response (UPR) are critical in decidualized cells.
- Dysregulation of these pathways is implicated in reproductive failures such as recurrent pregnancy loss (RPL) and recurrent implantation failure (RIF).
Purpose of the Study:
- To investigate the role of microRNAs (miRNAs) in regulating ERS and UPR in decidualized cells.
- To explore the association between miRNA expression and reproductive failures.
Main Methods:
- In vitro decidualization of endometrial stromal cells (St-T1b) with 8-Br-cAMP.
- Induction of ERS using thapsigargin.
- Quantitative PCR for ERS sensors, UPR markers, and miRNA expression.
- In-silico analysis to identify potential miRNA regulators.
- In-situ hybridization on endometrial biopsies from patients with RPL and RIF.
Main Results:
- Decidualization increased expression of ERS sensors (ATF6α, PERK, IRE1α) and UPR marker (CHOP).
- Inflammatory markers (TXNIP, NLRP3, IL-1β) were upregulated during decidualization.
- miR-17-5p, miR-21-5p, and miR-193b-3p were identified as potential regulators of ERS/UPR and inflammation.
- These miRNAs showed decreased expression in thapsigargin-treated non-decidualized cells.
- Differential expression of these miRNAs was observed in stromal cells from patients with RPL and RIF, with lower expression in RPL.
Conclusions:
- Decidualization involves ERS/UPR and inflammatory processes potentially modulated by specific miRNAs.
- Altered expression of miR-17-5p, miR-21-5p, and miR-193b-3p in stromal cells from patients with reproductive failure indicates a role in regulating ERS/UPR pathways.
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