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JoVE Journal
Neuroscience
Bioimpresión tridimensional de cocultivos de neuronas-astrocitos derivados de iPSC humanos para a...
Bioimpresión tridimensional de cocultivos de neuronas-astrocitos derivados de iPSC humanos para a...
JoVE Journal
Neuroscience
This content is Free Access.
JoVE Journal Neuroscience
Three-Dimensional Bioprinting of Human iPSC-Derived Neuron-Astrocyte Cocultures for Screening Applications

Bioimpresión tridimensional de cocultivos de neuronas-astrocitos derivados de iPSC humanos para aplicaciones de cribado

Full Text
5,610 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.

Aquí, presentamos un protocolo para producir cocultivos bioimpresos en 3D de neuronas y astrocitos derivados de iPSC. Este modelo de cocultivo, generado dentro de un andamio de hidrogel en formatos de 96 o 384 pocillos, demuestra una alta viabilidad post-impresión y crecimiento de neuritas en 7 días y muestra la expresión de marcadores de madurez para ambos tipos de células.

El modelado celular 3D es un campo novedoso que se ha expandido exponencialmente en la última década. Se ha demostrado que estos modelos facilitan el crecimiento neuronal y representan con mayor precisión los fenotipos de la enfermedad. Sin embargo, creemos que hay un cambio hacia la creación de estos modelos de mayor rendimiento y la necesidad de adoptar la automatización dentro del desarrollo.

Los métodos tradicionales de desarrollo de cultivos 3D pueden ser laboriosos y llevar mucho tiempo establecerlos, pero la bioimpresión 3D es una tecnología que se puede aplicar para ampliar estos procesos de desarrollo. Esta tecnología permite crear cientos de modelos idénticos de manera eficiente y sin errores humanos. Este protocolo desarrolla cultivos complejos porque las células neuronales se cultivan en 3D en matrices de hidrogel biológicamente activas.

Pero lo más importante es que este protocolo prioriza la velocidad y la comodidad en el desarrollo de modelos, que pueden faltar en este campo y pueden dificultar la implementación en la industria. Este protocolo define un método para establecer muchos cocultivos 3D de manera muy eficiente con una participación limitada de los usuarios. Esperamos que esto elimine las barreras para el uso de modelos complejos de cultivo celular dentro de ensayos de alto rendimiento y facilite una mayor investigación del efecto del cultivo 3D en los tipos de células neuronales.

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Bioimpresión tridimensional Cocultivos neurona-astrocitos derivados de IPSC humana Aplicaciones de cribado Modelo celular Cribado de fármacos Rendimiento Homogeneidad Tiempo de desarrollo Modelos 3D Bioimpresión Modelos de cocultivo Células madre pluripotentes inducidas (iPSC) Neuronas glutamatérgicas Astrocitos Matriz de hidrogel Péptidos bioactivos Proteínas de matriz extracelular (MEC) Rigidez fisiológica Viabilidad Relación astrocito-neurona Marcadores de tipo de células neurales maduras Ensayos de crecimiento de neuritas

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