Establishing 3D Endometrial Organoids from the Mouse Uterus

Suni Tang; Sydney E. Parks; Zian Liao; Dominique I. Cope; Sarah E. Blutt; Diana Monsivais

doi:10.3791/64448

إنشاء عضويات بطانة الرحم 3D من رحم الفأر

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Establishing 3D Endometrial Organoids from the Mouse Uterus

إنشاء عضويات بطانة الرحم 3D من رحم الفأر

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06:24 min

January 6, 2023

DOI: 10.3791/64448-v

Suni Tang^1,2, Sydney E. Parks^1,2, Zian Liao^1,2, Dominique I. Cope^1,2, Sarah E. Blutt³, Diana Monsivais^1,2

¹Department of Pathology & Immunology,Baylor College of Medicine, ²Center for Drug Discovery,Baylor College of Medicine, ³Departments of Molecular Virology and Microbiology and Medicine,Baylor College of Medicine

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Overview

This protocol provides a step-by-step methodology for isolating highly pure uterine epithelium from the mouse uterus to culture endometrial organoids. The technique utilizes both enzymatic and mechanical dissociation, allowing for effective establishment, treatment, and analysis of the organoids.

Key Study Components

Research Area

Endometrial biology
Organoid culture
Mouse models

Background

Importance of studying endometrial epithelium
Protocols for organoid establishment
Applications in understanding uterine conditions

Methods Used

Enzymatic and mechanical dissociation methods
Mouse uterus as the biological system
Phase contrast imaging, immunostaining, and qPCR

Main Results

Successful isolation of endometrial epithelium with minimal contamination
Organoid cultures formed within days
Demonstrated gene expression changes in response to estradiol treatment

Conclusions

The method allows for the study of endometrial signaling and renewal
Potential applications for treating endometrial-related diseases

Frequently Asked Questions

What is an endometrial organoid?

Endometrial organoids are 3D structures derived from endometrial epithelium that mimic the uterine environment for research.

Why use mouse models for this research?

Mouse models provide a controlled environment to study mammalian biology and disease processes.

How does estradiol influence epithelial organoids?

Estradiol has been shown to significantly increase the expression of specific genes related to endometrial function.

What technologies are utilized in this protocol?

This protocol employs enzymatic dissociation, microscopy, and quantitative PCR to analyze the organoids.

What are the potential applications of this research?

It has implications for understanding diseases like endometriosis, pregnancy loss, and cancer.

What is the importance of clear epithelial separation?

Clear separation ensures the integrity and purity of the isolated organoids, which is crucial for accurate experimental results.

How long does it take to establish organoid cultures?

Endometrial organoids can be established within three to four days following the protocol.

يصف هذا البروتوكول منهجيات إنشاء عضويات ظهارية بطانة الرحم للفأر للتعبير الجيني والتحليلات النسيجية.

هذا البروتوكول مهم لأنه يظهر منهجية خطوة بخطوة لعزل ظهارة الرحم النقية للغاية من رحم الفأر لثقافة عضويات بطانة الرحم. تستخدم هذه الطريقة بكفاءة التفكك الأنزيمي والميكانيكي لعزل ظهارة الرحم. تم تبسيط التقنيات لإنشاء ومعالجة وتحليل عضويات ظهارة بطانة الرحم المشتقة من الظهارة المعزولة.

ابدأ بإذابة مصفوفة الجل على الثلج لمدة ساعة إلى ساعتين تقريبا قبل الاستخدام. ثم قم بتشريح الفأر الرحيم عن طريق إجراء شق خط الوسط على البطن باستخدام المقص ، وتقشير الجلد برفق لفضح الطبقة البريتونية الأساسية. حدد موقع قرون الرحم عن طريق تحريك ضمادات الدهون في البطن جانبا برفق وأمسك قرون الرحم عند تقاطع عنق الرحم قبل قطعها على طول الدهون المساريقي.

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