Isolation and Characterization of Patient-derived Pancreatic Ductal Adenocarcinoma Organoid Models

Herv&#233; Tiriac; Randall French; Andrew  M. Lowy

doi:10.3791/60364

JoVE Journal
Cancer Research

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JoVE Journal Cancer Research

Isolation and Characterization of Patient-derived Pancreatic Ductal Adenocarcinoma Organoid Models

患者由来膵管腺癌オルガノイドモデルの単離と特徴付け

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08:43 min

January 14, 2020

DOI: 10.3791/60364-v

Hervé Tiriac^1,2, Randall French^1,2, Andrew M. Lowy^1,2

¹Moores Cancer Center,University of California San Diego, ²Department of Surgery, Division of Surgical Oncology,University of California San Diego

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Overview

Patient-derived organoid cultures of pancreatic ductal adenocarcinoma provide a 3-dimensional model that accurately represents tumor cell compartments, facilitating translational research. This method allows for rapid establishment and propagation of organoids, enabling relevant biological assays.

Key Study Components

Area of Science

Neuroscience
Oncology
Translational Research

Background

Patient-derived organoid models reflect the genetic and phenotypic heterogeneity of pancreatic cancer.
Organoids can be isolated from limited cancer samples with high success rates.
They can be rapidly expanded in vitro for further analysis.
Studying organoid responses to therapies may reveal mechanisms of drug resistance.

Purpose of Study

To establish a reliable method for creating organoid cultures from pancreatic cancer patients.
To enable the study of drug responses in a controlled environment.
To contribute to the development of effective treatment strategies for pancreatic cancer.

Methods Used

Isolation of organoids from patient-derived samples.
Propagation of organoids in a 12-well plate.
Use of basement membrane extract for organoid culture.
Assays to evaluate drug responses in organoid models.

Main Results

Successful establishment of organoid cultures from pancreatic cancer samples.
Organoids demonstrated the ability to reflect tumor heterogeneity.
Responses to cytotoxic drugs were analyzed to understand resistance mechanisms.
Potential for organoids to inform treatment strategies was highlighted.

Conclusions

Patient-derived organoids are a valuable tool for studying pancreatic cancer.
The method allows for rapid and effective analysis of tumor biology.
Organoids can aid in the identification of new therapeutic approaches.

Frequently Asked Questions

What are patient-derived organoids?

Patient-derived organoids are 3D cell cultures derived from patient tumors that mimic the original tumor's characteristics.

How are organoids established?

Organoids are established by isolating cells from tumor samples and culturing them in a specialized medium.

What is the significance of using organoids in cancer research?

Organoids allow researchers to study the genetic and phenotypic diversity of tumors and test drug responses in a controlled setting.

Can organoids be used for drug testing?

Yes, organoids can be used to evaluate the effectiveness of various drugs and understand mechanisms of resistance.

What advantages do organoids have over traditional cell cultures?

Organoids better replicate the tumor microenvironment and maintain the heterogeneity of the original tumor.

How quickly can organoids be propagated?

Organoids can be expanded rapidly in vitro, allowing for timely analysis and experimentation.

膵管腺癌の患者由来オルガノイド培養は、高忠実度で上皮腫瘍細胞コンパートメントを表す急速に確立された3次元モデルであり、この致死性悪性腫瘍に対する翻訳研究を可能にする。ここでは、オルガノイドを確立し、伝播する方法を詳細に提供するとともに、これらのモデルを用いて関連する生物学的アッセイを行う方法を提供する。

患者由来のオルガノイドモデルは、この疾患の遺伝的および表現型異質性を反映した膵臓癌を研究するための強力かつ迅速なツールであるため、この方法は重要である。この技術の主な利点は、オルガノイドは限られた量の癌から高い成功率で単離することができ、下流の分析のためにインビトロで急速に拡大することができるということです。細胞毒性薬に対するオルガノイドの反応を研究することは、腫瘍が治療に耐性になる理由を理解し、より効果的な治療戦略を定義するのに役立つかもしれません。

この手順を開始するには、調製した12ウェルプレートをインキュベーターから取り出します。無菌セルリフターを使用して、底面を削らないようにしながら、完全な媒体に浮遊するように、地下膜エキスドームをそっと持ち上げます。P1000ピペットを使用して、BMEドームとメディアを慎重に15ミリリットルの円錐チューブに移します。

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