Initial 3D Cell Cluster Control in a Hybrid Gel Cube Device for Repeatable Pattern Formations

Masaya Hagiwara; Rina Nobata; Tomohiro Kawahara

doi:10.3791/59214

Celular 3D inicial grupo Control en un dispositivo híbrido Gel cubo para formaciones de patrón re...

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Bioengineering

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JoVE Journal Bioengineering

Initial 3D Cell Cluster Control in a Hybrid Gel Cube Device for Repeatable Pattern Formations

Celular 3D inicial grupo Control en un dispositivo híbrido Gel cubo para formaciones de patrón repetible

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05:22 min

March 21, 2019

DOI: 10.3791/59214-v

Masaya Hagiwara¹, Rina Nobata², Tomohiro Kawahara³

¹NanoSquare Research Institute,Osaka Prefecture University, ²Department of Bioscience and Informatics,Osaka Prefecture University, ³Department of Biological Functions Engineering,Kyushu Institute of Technology

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Overview

This protocol presents a method for controlling the initial shape of cell clusters in a 3D extracellular matrix, facilitating repeatable pattern formation. Utilizing a cubic device with two different hydrogels, the technique allows for multi-directional imaging in tissue pattern formation.

Key Study Components

Area of Science

Neuroscience
Cell Biology
Tissue Engineering

Background

3D cell culture models are essential for studying tissue development.
Controlling initial culture conditions can enhance experimental reliability.
Hydrogels are commonly used in tissue engineering for their biocompatibility.
Multi-directional imaging provides comprehensive insights into tissue formation.

Purpose of Study

To develop a reliable method for 3D cell culture without specialized equipment.
To achieve consistent pattern formation in cell clusters.
To utilize a cubic device for enhanced imaging capabilities.

Methods Used

Preparation of a polycarbonate cubic frame for cell culture.
Application of preheated agarose to create a flat surface.
Sequential filling of the cubic frame with agarose to form walls.
Utilization of tweezers for precise manipulation of the frame.

Main Results

Successful creation of a stable 3D cell culture environment.
Repeatable pattern formation observed in cell clusters.
Effective use of hydrogels for multi-directional imaging.
Enhanced reliability of experimental outcomes in tissue studies.

Conclusions

The method provides a straightforward approach to 3D cell culture.
Controlling initial conditions is crucial for reproducibility.
This technique can be applied in various tissue engineering applications.

Frequently Asked Questions

What are the advantages of using a cubic device?

The cubic device allows for controlled initial conditions and facilitates multi-directional imaging.

Is special equipment required for this protocol?

No, the protocol can be performed without any special equipment.

What materials are used in this method?

The method primarily uses polycarbonate frames and agarose hydrogels.

How does this method enhance reliability in experiments?

By controlling the initial culture conditions, the method ensures consistent outcomes in 3D cultures.

Can this technique be applied to other types of cells?

Yes, the technique can be adapted for various cell types in tissue engineering.

What is the significance of multi-directional imaging?

Multi-directional imaging provides a comprehensive view of tissue development and pattern formation.

Presentamos un procedimiento para el control de la forma de racimo de célula inicial en una matriz extracelular 3D para obtener una formación de patrón repetible. Un dispositivo cúbico que contiene dos diferentes hidrogeles se emplea para lograr la proyección de imagen multi-direccional para la formación de patrones de tejido.

Este protocolo utiliza un método simple para mejorar la fiabilidad de un modelo de cultivo celular 3D sin el uso de ningún equipo especial. La principal ventaja de esta técnica es que podemos llevar a cabo el experimento de cultivo 3D repetible controlando la condición de cultivo inicial dentro del dispositivo cúbico. Después de preparar un marco cúbico de policarbonato de cinco por cinco milímetros, coloque el marco en una diapositiva preenfriada y una caja de hielo y utilice una pipeta para agregar 12 microlitros de 1.5% de aguantó previamente desde la cara superior del marco cúbico a la superficie inferior.

Esparce la agarosa para obtener una superficie plana y permitir que el polímero se cure. Utilice pinzas para deslizar el marco hasta el borde del cristal y gire el marco de modo que el lado abierto esté mirando hacia abajo antes de volver a colocar el marco en la diapositiva. Después de llenar las siguientes dos superficies del marco con más agarosa, como se acaba de demostrar, deje caer la agarosa en un recipiente de una cara abierta para formar una pared de agarosa en los lados cuarto y quinto.

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