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Neuroscience
Bioimpressão Tridimensional de Coculturas Neurônio-Astrócitos Derivadas de iPSC Humanas para Apli...
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Three-Dimensional Bioprinting of Human iPSC-Derived Neuron-Astrocyte Cocultures for Screening App...
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Neuroscience
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JoVE Journal Neuroscience
Three-Dimensional Bioprinting of Human iPSC-Derived Neuron-Astrocyte Cocultures for Screening Applications

Bioimpressão Tridimensional de Coculturas Neurônio-Astrócitos Derivadas de iPSC Humanas para Aplicações de Triagem

Full Text
5,583 Views
08:03 min
September 29, 2023

DOI: 10.3791/65856-v

Chloe Ann Whitehouse1, Yufang He2, Janet Brownlees1, Nicola Corbett1

1MSD Research Laboratories, London, UK, 2Merck & Co., Inc., Rahway, NJ, USA

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Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This study presents a protocol for the efficient production of 3D-bioprinted cocultures of iPSC-derived neurons and astrocytes within hydrogel scaffolds. The developed model operates in 96- or 384-well formats and demonstrates high post-print viability and neurite outgrowth within seven days, while expressing maturity markers for both cell types. This approach aims to enhance the throughput and automation of 3D cell culture systems.

Key Study Components

Area of Science

  • Neuroscience
  • Cell Biology
  • Biotechnology

Background

  • 3D cell modeling has rapidly advanced, enabling more accurate representations of disease phenotypes.
  • Traditional methods for 3D culture development are often labor-intensive and time-consuming.
  • 3D bioprinting offers a solution by automating and scaling the development of complex cultures.
  • This technology can produce numerous identical models rapidly, minimizing human error in the process.

Purpose of Study

  • To create a high-throughput protocol for establishing 3D cocultures of neural cells.
  • To improve the speed and convenience of model development in neuroscience research.
  • To facilitate further investigation into the effects of 3D culture environments on neural cell types.

Methods Used

  • The platform utilizes 3D bioprinting to create cocultures within hydrogel matrices.
  • The biological model includes iPSC-derived neurons and astrocytes, grown in a controlled 3D environment.
  • This protocol emphasizes efficient coculture establishment with minimal user intervention needed.
  • Key timelines involve assessing cell viability and neurite outgrowth within a seven-day period.
  • Hydrogel scaffolds are employed to support the structural integrity and functionality of the cells.

Main Results

  • The coculture model exhibited high post-print cell viability and significant neurite outgrowth within seven days.
  • Both cell types demonstrated the expression of maturity markers, indicating successful development.
  • Rapid model creation supports more streamlined research applications and further testing.
  • Conclusions highlight the potential of this protocol to overcome existing barriers in complex cell culture models.

Conclusions

  • The study demonstrates a novel method for developing scalable 3D coculture systems in neuroscience research.
  • This advancement may significantly impact the future of high-throughput assays in neural research.
  • Insights gained from using 3D cultures could enhance understanding of neural mechanisms and plasticity.

Frequently Asked Questions

What are the advantages of using 3D bioprinting for cocultures?
3D bioprinting allows for high-throughput production of complex cellular models with improved reproducibility and reduced manual interventions, enhancing experimental efficiency.
How are the biological models implemented in this study?
The biological model consists of iPSC-derived neurons and astrocytes grown within a hydrogel scaffold to support 3D cellular interactions and promote differentiation.
What types of data can be obtained from this model?
The model allows for assessments of cell viability, neurite outgrowth, and maturity marker expression, providing insights into neural development and function.
How can this method be adapted for various research needs?
The protocol's scalability and automation may be adjusted for different cell types or experimental conditions, making it versatile for diverse neuroscience applications.
What key limitations should be considered?
While the protocol streamlines model development, careful optimization may still be required for specific experimental contexts, particularly regarding hydrogel formulation and cell sourcing.
How does this research contribute to understanding neural mechanisms?
By providing a robust 3D culture system, the study offers a platform for exploring cellular interactions and plasticity, enhancing insights into neural behavior and disease models.

Aqui, apresentamos um protocolo para produzir coculturas bioimpressas em 3D de neurônios e astrócitos derivados de iPSC. Este modelo de cocultura, gerado dentro de um arcabouço de hidrogel em formatos de 96 ou 384 poços, demonstra alta viabilidade pós-impressão e crescimento de neuritos em 7 dias e mostra a expressão de marcadores de maturidade para ambos os tipos celulares.

A modelagem de células 3D é um campo novo que se expandiu exponencialmente na última década. Esses modelos têm demonstrado facilitar o crescimento neuronal e representar com mais precisão os fenótipos da doença. No entanto, acreditamos que há uma mudança no sentido de tornar esses modelos de maior rendimento e uma necessidade de adotar a automação dentro do desenvolvimento.

Os métodos tradicionais de desenvolvimento de culturas 3D podem ser trabalhosos e demorados para serem estabelecidos, mas a bioimpressão 3D é uma tecnologia que pode ser aplicada para escalar esses processos de desenvolvimento. Essa tecnologia permite que centenas de modelos idênticos sejam criados de forma eficiente e sem erro humano. Este protocolo desenvolve culturas complexas porque as células neurais são cultivadas em 3D em matrizes hidrogéis biologicamente ativas.

Mas, criticamente, esse protocolo prioriza a velocidade e a conveniência no desenvolvimento do modelo, o que pode faltar nesse campo e dificultar a implementação na indústria. Este protocolo define um método para estabelecer muitas coculturas 3D de forma muito eficiente com entrada limitada dos usuários. Esperamos que isso remova as barreiras ao uso de modelos complexos de cultura de células em ensaios de alto rendimento e facilite a investigação adicional do efeito da cultura 3D em tipos de células neurais.

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