Tumorsphere Derivation and Treatment from Primary Tumor Cells Isolated from Mouse Rhabdomyosarcomas

Francesca Boscolo Sesillo; Alessandra Sacco

doi:10.3791/59897

Derivação e tratamento de tumorsphere de células tumorais primárias isoladas de rabdomiossarcoma ...

JoVE Journal
Genetics

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JoVE Journal Genetics

Tumorsphere Derivation and Treatment from Primary Tumor Cells Isolated from Mouse Rhabdomyosarcomas

Derivação e tratamento de tumorsphere de células tumorais primárias isoladas de rabdomiossarcoma de camundongo

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09:21 min

September 13, 2019

DOI: 10.3791/59897-v

Francesca Boscolo Sesillo^1,2,3, Alessandra Sacco²

¹Graduate School of Biomedical Sciences,Sanford Burnham Prebys Medical Discovery Institute, ²Development, Aging and Regeneration Program,Sanford Burnham Prebys Medical Discovery Institute, ³Department of Obstetrics, Gynecology, and Reproductive Sciences, Division of Female Pelvic Medicine and Reconstructive Surgery,University California San Diego

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Overview

This protocol describes a reproducible method for the isolation of mouse rhabdomyosarcoma primary cells, tumorsphere formation, and treatment, as well as allograft transplantation starting from tumorsphere cultures. The tumorsphere formation assay is highlighted for its ability to mimic the tumor environment and facilitate drug screening studies.

Key Study Components

Area of Science

Neuroscience
Oncology
Cell Biology

Background

Rhabdomyosarcoma is a rare soft-tissue sarcoma.
Characterized by specific tissue morphology and myogenic markers.
Challenges exist in identifying the cell of origin due to tumor heterogeneity.
Freshly harvested tumor tissues from mice are used for study.

Purpose of Study

To identify the cell of origin of rhabdomyosarcomas.
To establish a reliable method for tumorsphere formation.
To enable drug screening using tumorsphere models.

Methods Used

Isolation of primary cells from mouse rhabdomyosarcoma.
Formation of tumorspheres from isolated cells.
Treatment of tumorspheres with various agents.
Allograft transplantation of tumorspheres into mice.

Main Results

Successful isolation and culture of rhabdomyosarcoma primary cells.
Formation of tumorspheres that mimic the tumor microenvironment.
Potential for drug screening applications using tumorsphere models.
Method applicable to other tumor types with optimization.

Conclusions

The tumorsphere formation assay is a valuable tool for studying rhabdomyosarcoma.
This method can enhance our understanding of tumor biology.
It offers a platform for testing therapeutic agents.

Frequently Asked Questions

What is rhabdomyosarcoma?

Rhabdomyosarcoma is a rare form of soft-tissue sarcoma characterized by specific tissue morphology and myogenic markers.

Why is tumorsphere formation important?

Tumorsphere formation mimics the tumor environment and allows for the expansion and enrichment of specific cell populations.

How are primary cells isolated?

Primary cells are isolated from freshly harvested tumor tissues from mice.

What are the applications of this method?

This method can be used for drug screening studies and may be applicable to other tumor types with optimization.

What are the main advantages of the tumorsphere assay?

The assay relies on cellular properties present in tumor-originating cells and mimics the tumor microenvironment.

Can this method be optimized for other tumors?

Yes, the method may require optimization of culture conditions for different tumor types.

Este protocolo descreve um método reprodutível para a isolação de pilhas preliminares do rabdomiossarcoma do rato, da formação e do tratamento do tumorsphere, e da transplantação do aloenxerto que partem das culturas dos tumorspheres.

Rabdomiossarcoma é uma forma rara de sarcomas de tecido mole caracterizada histologicamente pela morfologia tecidual e pela expressão de marcadores miogênicos. No entanto, dada a existência de diferentes subtipos desses tumores e a heterogeneidade dos estímulos que contribuem para sua formação, a identificação da célula de origem tem sido muito desafiadora. Aqui descrevemos métodos reprodutíveis e confiáveis para a identificação da célula de origem dos rabdomiossarcomas a partir da colheita recente de tecidos tumorais de camundongos e este ensaio é o ensaio de formação tumorais.

As principais vantagens do ensaio de formação tumorais é que ele se baseia em propriedades celulares que são conhecidas por estarem presentes em células originárias de tumores e também pode ser usada para expansão e enriquecimento da população celular específica. Além disso, sua estrutura esferoide imita o ambiente tumoral, podendo ser usados como estudos de rastreamento de drogas. Este método tem o potencial de ser aplicado a outros tipos de tumores porque não requer conhecimento prévio de marcadores moleculares, mas pode exigir otimização das condições culturais.

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