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Published on: June 21, 2024
Development of an in vitro model for the analysis of bovine endometrium using simple techniques
Keisuke Yamauchi1, Nobuhiko Yamauchi, Kazuki Yamagami
1Department of Animal and Marine Bioresource Sciences, Graduate School Kyushu University, Fukuoka, Japan.
Insights
Researchers developed an in vitro bovine endometrium model using spheroids and explants. This model effectively mimics endometrial cell structure and receptor expression, offering a new platform for studying bovine endometrial function.
Area of Science:
- Veterinary Science
- Reproductive Biology
- Cell Biology
Background:
- The bovine endometrium plays a crucial role in reproductive success.
- Accurate in vitro models are essential for studying endometrial function and disease.
- Existing models may not fully replicate the complex cellular architecture and molecular responses of the native endometrium.
Purpose of the Study:
- To develop and validate an in vitro model for analyzing the bovine endometrium.
- To compare the structural and functional characteristics of spheroid and explant models.
- To assess the expression of key receptors and enzymes in response to hormonal stimuli.
Main Methods:
- Development of homo- and hetero-spheroids and cultured explants from bovine endometrial tissue.
- Immunofluorescent staining for cell localization (epithelial and stromal cells).
- Gelatin zymography for matrix metalloproteinase (MMP) activity.
- Quantitative real-time PCR for messenger RNA (mRNA) expression of progesterone receptor (PR), estrogen receptor alpha (ERα), interferon receptors (IFNAR1, IFNAR2), oxytocin receptor (OTR), and hepatocyte growth factor (HGF).
- Hormonal treatments (E2, P4) to assess receptor-mediated responses.
Main Results:
- Both spheroid and explant models showed similar localization of endometrial epithelial and stromal cells.
- Spheroids did not express MMPs after 4 days, while explants showed strong MMP expression up to 7 days.
- Expression of PR, ERα, IFNAR1, and IFNAR2 mRNA was detected in both spheroid types.
- Hormonal treatments (E2, E2+P4) significantly increased OTR mRNA in spheroids and PR/OTR mRNA in explants.
- HGF mRNA expression was elevated in P4-treated explants at 10 days.
Conclusions:
- The developed in vitro model, utilizing spheroids and explants, effectively mimics bovine endometrial cell structure and key receptor expression.
- The model demonstrates differential expression of MMPs between spheroids and explants, providing insights into tissue remodeling.
- This model serves as a valuable platform for future research into bovine endometrial physiology and pathology, including responses to hormonal signaling.
Abstract:
This study aimed to develop an in vitro model for the analysis of the bovine endometrium. Immunofluorescent staining revealed that the hetero-spheroids and the cultured explants showed almost similar structure in the localization of bovine endometrial epithelial cells and endometrial stromal cells, except the glandular-like structure of the epithelial cells inside the explants. Gelatin zymography revealed that the hetero-spheroids did not express matrix metalloproteinases (MMPs) after 4 days of culture, but strong MMP expressions were observed in the cultured explants until 7 days of culture. Additionally, expression of progesterone receptor (PR), estrogen receptor alpha (ERα), type I interferon receptor 1 (IFNAR1) and 2 (IFNAR2) messenger RNA was observed both in the homo- and hetero-spheroids. The expression of oxytocin receptor (OTR) mRNA in E2 and E2+P4 (1,3,5(10)-Estratrien-3, 17β-diol + 4-Pregnen-3, 20-dinone) treated groups were significantly (P < 0.05) higher than that of the control group of spheroids. In case of cultured explants, the expression of PR and OTR mRNA were significantly (P < 0.05) higher in E2 treated groups compared to the control groups. Hepatocyte growth factor (HGF) mRNA expression was also higher in P4 treated groups at 10 days in culture (P < 0.05). In a nutshell, the in vitro model developed in this study for the analysis of the endometrium may provide a new platform for extensive research on bovine endometrial function.
