Dissociation of Human and Mouse Tumor Tissue Samples for Single-cell RNA Sequencing

Jacy Fang; Isabella Salinas; Sarah San Vicente; Caroline Zielinski; Moshe Sade-Feldman

doi:10.3791/66766

Dissociação de amostras de tecido tumoral humano e de camundongo para sequenciamento de RNA de cé...

JoVE Journal
Cancer Research

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JoVE Journal Cancer Research

Dissociation of Human and Mouse Tumor Tissue Samples for Single-cell RNA Sequencing

Dissociação de amostras de tecido tumoral humano e de camundongo para sequenciamento de RNA de célula única

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05:58 min

August 16, 2024

DOI: 10.3791/66766-v

Jacy Fang*^1,2, Isabella Salinas*^1,2, Sarah San Vicente*^1,2, Caroline Zielinski*^1,2, Moshe Sade-Feldman^1,2,3

¹Krantz Family Center for Cancer Research, Department of Medicine,Massachusetts General Hospital, ²Broad Institute of MIT and Harvard, ³Department of Medicine,Harvard Medical School

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Overview

This protocol outlines the procedure for rapidly dissociating human and mouse tumor samples for single-cell RNA sequencing. This technique is crucial for understanding the immune response against tumors and identifying predictors of therapy resistance.

Key Study Components

Area of Science

Neuroscience
Immunology
Single-cell analysis

Background

Understanding mechanisms of response or resistance to immunotherapies.
Importance of real-time processing and analysis of patient samples.
Development of single-cell RNA sequencing techniques.
Need to investigate cellular compartments within tumors.

Purpose of Study

To study the immune response against tumors.
To identify predictors of therapy success.
To overcome resistance to standard immunotherapies.

Methods Used

Single-cell RNA sequencing
Rapid dissociation of tumor samples
Real-time analysis of molecular features
Investigation of tumor ecosystems

Main Results

Insights into the immune response mechanisms.
Identification of key immunological determinants.
Enhanced understanding of tumor microenvironments.
Potential targets for overcoming therapy resistance.

Conclusions

Single-cell RNA sequencing is vital for tumor analysis.
Real-time processing aids in understanding complex tumors.
Future research can focus on identified predictors of therapy.

Frequently Asked Questions

What is single-cell RNA sequencing?

Single-cell RNA sequencing is a technique that allows for the analysis of gene expression at the individual cell level, providing insights into cellular heterogeneity.

Why is rapid dissociation of tumor samples important?

Rapid dissociation preserves the integrity of the cells and allows for timely analysis of their molecular features, which is crucial for understanding tumor biology.

How does this study contribute to immunotherapy research?

This study helps identify predictors of immune response and resistance, which can inform the development of more effective immunotherapy strategies.

What are the challenges in studying human immunity?

Challenges include the complexity of human immune responses and the need for real-time analysis of patient-derived samples.

What advancements have been made in tumor analysis?

Recent advancements include the integration of single-cell RNA sequencing techniques that allow for unbiased analysis of tumor ecosystems.

What is the significance of understanding tumor ecosystems?

Understanding tumor ecosystems is crucial for identifying how different cell types interact and contribute to tumor growth and response to therapies.

Este protocolo descreve o procedimento para dissociar rapidamente amostras de tumores humanos e de camundongos para sequenciamento de RNA de célula única.

O escopo de nossa pesquisa é estudar os mecanismos de resposta ou resistência em imunoterapias padrão de tratamento. Nosso laboratório está tentando entender o que impulsiona uma resposta imune bem-sucedida contra tumores e se podemos identificar esses preditores de terapia para encontrar alvos que possam superar a resistência. O maior avanço nos últimos anos foi o desenvolvimento e integração de técnicas de sequenciamento de RNA de célula única.

Eles nos permitem, em tempo real, estudar sistematicamente as características moleculares em células individuais de maneira imparcial, o que é útil na dissecação desses ecossistemas tumorais complexos e em evolução. Embora muitos modelos pré-clínicos estejam sendo usados para estudar os mecanismos de resposta à imunoterapia, ainda não conhecemos os determinantes imunológicos que são importantes na imunidade humana. Processar e analisar amostras de pacientes em tempo real é crucial para nossa capacidade de investigar todos os compartimentos celulares dentro de um determinado tumor.

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