Two-Photon Polymerization 3D-Printing of Micro-scale Neuronal Cell Culture Devices

Ali Hosseini; Giovanni Noselli; Michele Giugliano

doi:10.3791/66142

Polimerização de dois fótons Impressão 3D de dispositivos de cultura de células neuronais em micr...

JoVE Journal
Neuroscience
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JoVE Journal Neuroscience

Two-Photon Polymerization 3D-Printing of Micro-scale Neuronal Cell Culture Devices

Polimerização de dois fótons Impressão 3D de dispositivos de cultura de células neuronais em microescala

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07:38 min

June 7, 2024

DOI: 10.3791/66142-v

Ali Hosseini¹, Giovanni Noselli², Michele Giugliano¹

¹Neuroscience Department,International School for Advanced Studies, ²Mathematics Department,International School for Advanced Studies

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Overview

This study explores the use of 3D printing at the micrometer scale to create polymeric devices for neuronal cell cultures, enabling the design of modular neural networks. By constraining structural connections between neurons through barriers and channels, the researchers investigated the functional consequences on neurite outgrowth using extracellular electrophysiology.

Key Study Components

Area of Science

Neuroscience
Cell Biology
3D Bioprinting

Background

The relationship between structure and function in neuronal microcircuits.
Utilizing dissociated neuron cell cultures for studying electrical activity and synaptic transmission.
Importance of rapidly fabricated devices for experimental research.
A focus on drug screening and pathological condition investigation.

Purpose of Study

To demonstrate rapid fabrication of microscale polymeric devices.
To facilitate the experimental study of modular neural networks in vitro.
To define the structure and control biological signal flow.

Methods Used

3D printing techniques to create PDMS-based devices for neuronal cultures.
Isolation and culturing of rat pup neuronal cells on microelectrode arrays (MEAs).
Extracellular electrophysiology to monitor neuronal activity.
Steps included design software for 3D components, UV curing, and device sterilization.

Main Results

Live imaging showed neurite outgrowth influenced by structural modifications.
Electrophysiological analysis indicated significant neuronal activity patterns in response to structural constraints.
Findings underscore the importance of physical structure in fostering cellular interactions.

Conclusions

The study demonstrates a novel method for creating tailored neuronal environments in vitro.
Encourages further exploration of engineered neural networks for research and clinical applications.
Implications for understanding neuronal mechanisms and disease modeling.

Frequently Asked Questions

What advantages does 3D printing offer for neuronal studies?

3D printing allows for precise control over the microenvironment of neuronal cultures, facilitating the fabrication of complex geometries that influence cell behavior and connectivity.

How are neuronal cultures implemented in this study?

Neuronal cultures are prepared by isolating cells from rat pup brains and seeding them onto microelectrode arrays integrated into the 3D printed devices.

What types of data are obtained from the electrophysiological recordings?

The recordings reveal patterns of neuronal activity and responses to structural constraints, providing insights into synaptic transmission and circuit dynamics.

Can this method be adapted for studying different neuronal types?

Yes, this method can be adapted for various neuronal types and experimental conditions, enhancing the versatility of in vitro neural network studies.

Are there any limitations in using 3D printed devices for neuronal cultures?

Limitations may include the scalability of device production and the necessity for careful calibration of printing parameters to ensure functionality in biological experiments.

A impressão 3D em escala micrométrica permite a prototipagem rápida de dispositivos poliméricos para culturas de células neuronais. Como prova de princípio, as conexões estruturais entre os neurônios foram restringidas pela criação de barreiras e canais que influenciam o crescimento dos neuritos, enquanto as consequências funcionais de tal manipulação foram observadas pela eletrofisiologia extracelular.

Estamos amplamente interessados na relação entre funções estruturais e no cérebro, particularmente ao nível dos micro-circuitos. Muitas vezes empregamos preparações experimentais reduzidas, como cultura de células de neurônios dissociados, porque elas são um modelo precioso para investigar a atividade elétrica, a transmissão sináptica e, em geral, o surgimento de estados coletivos. O método que propomos é ideal para fabricação rápida de dispositivos poliméricos em microescala.

Isso pode ser imediatamente usado para o estudo experimental de redes neurais modulares em um prato, e não requer um alto nível de conhecimento técnico em fotolitografia mole. Este método e nossa simples prova de conceito permitem o projeto de redes neuronais modulares onde podemos definir a estrutura, e controlar o fluxo de sinais biológicos. A triagem de drogas e a investigação de condições patológicas se beneficiam imediatamente dessas sofisticadas redes neuronais in vitro.

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