Generation of Organoids from Mouse Extrahepatic Bile Ducts

Junya Shiota; Nureen  H. Mohamad Zaki; Juanita  L. Merchant; Linda  C. Samuelson; Nataliya Razumilava

doi:10.3791/59544

Generazione dei Organoids da dotti biliari Extrahepatic Mouse

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Developmental Biology

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JoVE Journal Developmental Biology

Generation of Organoids from Mouse Extrahepatic Bile Ducts

Generazione dei Organoids da dotti biliari Extrahepatic Mouse

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09:13 min

April 23, 2019

DOI: 10.3791/59544-v

Junya Shiota¹, Nureen H. Mohamad Zaki¹, Juanita L. Merchant^1,2, Linda C. Samuelson^1,2, Nataliya Razumilava¹

¹Departments of Internal Medicine,The University of Michigan, ²Molecular & Integrative Physiology,The University of Michigan

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Overview

This protocol describes the production of a mouse extrahepatic bile duct 3-dimensional organoid system. These biliary organoids can be maintained in culture to study cholangiocyte biology and are composed of polarized epithelial cells.

Key Study Components

Area of Science

Neuroscience
Cell Biology
Organ Models

Background

Limited access to preclinical cholangiopathy models.
Challenges with pluripotent stem cell and liver-derived cholangiocyte organoid models.
Need for adult-tissue specific and reproducible models.
Importance of studying cholangiocyte biology.

Purpose of Study

To develop a model for studying cholangiocyte biology.
To provide a reductionist and cost-efficient method for organoid culture.
To facilitate research in laboratories lacking access to human tissues.

Methods Used

Isolation of intrahepatic bile duct from adult mice.
Maintaining organoids in culture.
Studying tissue regeneration and cell-to-cell interactions.
Demonstration of the procedure by a post-doc researcher.

Main Results

Successful production of biliary organoids.
Organoids express markers of progenitor and biliary cells.
Capability of culturing an unlimited number of cholangiocytes.
Potential for studying cholangiocyte biology and interactions.

Conclusions

The developed organoid system is a valuable tool for cholangiocyte research.
It addresses limitations of existing models.
Offers a reproducible method for studying cholangiopathies.

Frequently Asked Questions

What are biliary organoids?

Biliary organoids are 3D structures derived from the extrahepatic bile duct that can be cultured to study cholangiocyte biology.

Why is this model important?

It provides a specific and reproducible model for studying cholangiopathies, especially for labs without access to human tissues.

How are the organoids maintained?

The organoids can be maintained in culture, allowing for extensive study of their properties and behaviors.

Who demonstrated the procedure?

The procedure was demonstrated by Junya Shiota, a post-doc from the laboratory.

What are the advantages of this model?

The model is adult-tissue specific, reductionist, reproducible, and time and cost efficient.

What can be studied using these organoids?

Researchers can study tissue regeneration and cell-to-cell interactions in cholangiocytes.

Questo protocollo descrive la produzione di un sistema di 3-dimensionale organoid del mouse dei dotti biliari extrahepatic. Questi organoids biliare può essere mantenute in coltura per studiare biologia dei colangiociti. Organoids biliare esprimono marcatori di cellule progenitrici e cellule biliari e sono composte da cellule epiteliali polarizzate.

Questo sistema organoide del condotto biliare extraepatico murino può affrontare il limitato accesso ai modelli di colangiopatie precliniche e i limiti dei modelli organoidi di cellule staminali pluripotenti e corigiociti derivati dal fegato. Il nostro modello è specifico per i tessuti adulti, riduzionista, riproducibile ed efficiente in termini di tempo e costi. Sarà di particolare beneficio per i laboratori che non hanno accesso ai tessuti umani.

La nostra tecnica consente la coltivazione di un numero quasi illimitato di conlangiociti del condotto biliare extraepatico per studiare la rigenerazione dei tessuti e le interazioni cellula-cellula. A dimostrare la procedura sarà Junya Shiota, un post-doc del mio laboratorio. Per l'isolamento intraepatico del condotto biliare, posizionare il topo adulto in posizione supina e aprire la cavità addominale lungo la linea mediana.

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