Establishment and Characterization of Patient-derived Gastric Organoids from Biopsies of Benign Gastric Body and Antral Epithelium

Kole H. Buckley; Keely A. Beyries; Sandra Ryeom; Sam S. Yoon; Bryson W. Katona

doi:10.3791/66094

Estabelecimento e Caracterização de Organoides Gástricos Derivados de Pacientes a partir de Bióps...

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Establishment and Characterization of Patient-derived Gastric Organoids from Biopsies of Benign Gastric Body and Antral Epithelium

Estabelecimento e Caracterização de Organoides Gástricos Derivados de Pacientes a partir de Biópsias do Corpo Gástrico Benigno e Epitélio Antral

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08:45 min

January 26, 2024

DOI: 10.3791/66094-v

Kole H. Buckley¹, Keely A. Beyries¹, Sandra Ryeom², Sam S. Yoon², Bryson W. Katona¹

¹Division of Gastroenterology and Hepatology,University of Pennsylvania Perelman School of Medicine, ²Department of Surgery,Columbia University Irving Medical Center

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Overview

This research investigates the hereditary causes of gastric cancer, focusing on variants in the CDH1 and CTNNA1 genes. The study presents a reliable protocol for generating patient-derived gastric organoids from biopsy tissues obtained during upper endoscopy, demonstrating differences in growth rates between organoids derived from the antral and body regions of the stomach.

Key Study Components

Research Area

Hereditary causes of gastric cancer
Gastric organoid generation
Standardization of biological techniques

Background

Hereditary diffuse gastric cancer and genetic variants
Use of organoids in gastroenterology research
Standardized protocols for gastric organoid development

Methods Used

Isolation of single cells from gastric tissue biopsies
Patient-derived gastric organoids as the biological model
Cell digestion and seeding protocols in culture

Main Results

Organoids from antral biopsy tissue display larger size and growth rates compared to those from body tissue
Significant variations in organoid morphology across different patients
Standardized methods enable reliable growth comparison of organoids

Conclusions

The study establishes a reliable method for generating gastric organoids, aiding future research in gastric cancer.
Findings highlight the importance of tissue origin on organoid growth, which is crucial for experimental reproducibility.

Frequently Asked Questions

What are gastric organoids?

Gastric organoids are three-dimensional miniaturized organs derived from gastric tissue that mimic the structure and function of the stomach.

Why are organoids important in cancer research?

Organoids provide a realistic model for studying cancer biology and drug responses due to their ability to retain patient-specific features.

How are gastric organoids generated from patient biopsies?

They are generated using a single-cell digest method followed by seeding in a specialized matrix for culture.

What factors affect the growth of gastric organoids?

Tissue origin, culture conditions, and seeding density significantly influence organoid growth and morphology.

How long does it take for organoids to develop?

Gastric organoids typically become identifiable within 10 days post-seeding and reach significant growth by day 20.

Can these organoids be used for drug testing?

Yes, gastric organoids can be utilized for testing therapeutic responses, aiding in personalized medicine approaches.

What are the limitations of using organoids?

Although analogous to real tissues, organoids may lack some physiological complexities found in vivo, which could influence study outcomes.

Organoides gástricos derivados de pacientes encontram uso crescente em pesquisas, mas faltam protocolos formais para gerar organoides gástricos humanos a partir de digestores unicelulares com densidade de semeadura padronizada. Este protocolo apresenta um método detalhado para a criação confiável de organoides gástricos a partir de tecido de biópsia obtido durante a endoscopia digestiva alta.

Nossa pesquisa se concentra nas causas hereditárias do câncer gástrico. Em particular, estamos interessados no câncer gástrico difuso hereditário devido a variantes causadoras de doenças no gene CDH1 ou CTNNA1. Também investigamos o papel do BRCA1 e BRCA2 no risco de câncer gástrico e na carcinogênese.

Os organoides estão sendo cada vez mais utilizados em uma variedade de domínios de pesquisa e a gastroenterologia não é exceção, então com isso surge a necessidade de padronização dessas técnicas para gerar esses organoides, em particular organoides gástricos derivados do paciente. Nosso protocolo oferece um método passo-a-passo para gerar organoides gástricos derivados do paciente de forma confiável a partir de biópsias das regiões antral e corporal do estômago. Em particular, utilizamos uma abordagem de digestão de célula única para permitir a semeadura padronizada de células em um esforço para fazer comparações confiáveis entre organoides gerados de diferentes pacientes.

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