Generation of Microtumors Using 3D Human Biogel Culture System and Patient-derived Glioblastoma Cells for Kinomic Profiling and Drug Response Testing

Ashley N. Gilbert; Rachael S. Shevin; Joshua C. Anderson; Catherine P. Langford; Nicholas Eustace; G. Yancey Gillespie; Raj Singh; Christopher D. Willey

doi:10.3791/54026

Geração de Microtumors Usando 3D Biogel Humano Sistema de Cultura e células de glioblastoma deriv...

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JoVE Journal Medicine

Generation of Microtumors Using 3D Human Biogel Culture System and Patient-derived Glioblastoma Cells for Kinomic Profiling and Drug Response Testing

Geração de Microtumors Usando 3D Biogel Humano Sistema de Cultura e células de glioblastoma derivado do paciente para Kinomic Profiling e Testes resposta à droga

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09:24 min

June 9, 2016

DOI: 10.3791/54026-v

Ashley N. Gilbert¹, Rachael S. Shevin⁴, Joshua C. Anderson², Catherine P. Langford³, Nicholas Eustace², G. Yancey Gillespie³, Raj Singh⁴, Christopher D. Willey²

¹Biomedical Engineering,University of Alabama at Birmingham, ²Radiation Oncology,University of Alabama at Birmingham, ³Neurosurgery,University of Alabama at Birmingham, ⁴Vivo Biosciences, Inc.

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Overview

This study presents a patient-derived glioblastoma 3D microtumor culture system that enables high-throughput drug testing and molecular profiling. The method addresses critical questions in cancer therapy effectiveness across different patients.

Key Study Components

Area of Science

Neuroscience
Oncology
Drug Development

Background

Patient-derived xenografts are crucial for studying tumor biology.
3D culture systems mimic in vivo tumor environments.
Fragility of patient-derived cells complicates handling.
Visual demonstrations enhance understanding of complex methods.

Purpose of Study

To develop a realistic tumor culture system for therapy testing.
To investigate variability in therapy responses among patients.
To facilitate kinomic analysis alongside drug testing.

Methods Used

Creation of 3D microtumors from patient-derived xenografts.
High-throughput drug testing protocols.
Molecular profiling techniques, including kinomic analysis.
Visual demonstrations to aid in method learning.

Main Results

The 3D microtumor system successfully mimics patient tumor characteristics.
High-throughput testing revealed differential drug responses.
Kinomic analysis provided insights into signaling pathways.
Visual aids improved the learning curve for new users.

Conclusions

This method offers a promising platform for personalized cancer therapy research.
Understanding therapy variability can lead to better treatment strategies.
Further refinement of techniques will enhance reproducibility and accessibility.

Frequently Asked Questions

What are patient-derived xenografts?

Patient-derived xenografts are tumor cells taken from patients and implanted into immunocompromised mice for research purposes.

How does the 3D microtumor culture system work?

The system uses biogel to create a three-dimensional environment that mimics the in vivo tumor microenvironment for testing.

What is kinomic analysis?

Kinomic analysis involves studying the activity of kinases, which are enzymes that play key roles in cell signaling and cancer progression.

Why is visual demonstration important?

Visual demonstrations help researchers understand complex procedures that are difficult to grasp through text alone.

What are the advantages of using a 3D culture system?

3D culture systems provide a more accurate representation of tumor biology compared to traditional 2D cultures, leading to better drug response predictions.

Can this method be applied to other types of cancer?

While this study focuses on glioblastoma, the principles of the 3D culture system can potentially be adapted for other cancers.

Xenoenxertos derivados de pacientes de glioblastoma multiforme podem ser miniaturizados em microtumores vivos usando o sistema de cultura de biogel humano 3D. Este ensaio tumoral 3D semelhante ao in vivo é adequado para testes de resposta a medicamentos e perfis moleculares, incluindo análise cinômica.

O objetivo geral deste sistema de cultura de microtumor 3D de glioblastoma derivado do paciente é permitir testes de drogas de alto rendimento com um ensaio de perfil molecular correspondente, como testes cinômicos. Esse método pode ajudar a responder a perguntas-chave no campo do câncer, como: por que certas terapias funcionam bem no tumor de um paciente, mas não em outro? A principal vantagem dessa técnica é que ela fornece um sistema de cultura de tumor em miniatura, mas realista, adequado para testes de terapia.

Geralmente, os indivíduos novos neste método terão dificuldades porque manusear as células de xenoenxerto derivadas do paciente não é como lidar com células tumorais imortalizadas há muito estabelecidas. Isso se deve à sua fragilidade e sensibilidade às mudanças nas condições da cultura. A demonstração visual desse método é crítica, pois as etapas de geração do microtumor são difíceis de aprender apenas por meio da descrição, devido ao seu método único e complexo de produção.

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