A 3D Culture Method of Spheroids of Embryonic and Liver Zebrafish Cell Lines

Irisdoris Rodrigues de Souza; Andrezza Di Pietro Micali Canavez; Desiree Cigaran Schuck; Viviana Stephanie Costa Gagosian; Isisdoris Rodrigues de Souza; Nat&#225;lia de Albuquerque Vita; Edvaldo da Silva Trindade; Marta Margarete Cestari; Marcio Lorencini; Daniela Morais Leme

doi:10.3791/64859

A 3D Culture Method of Spheroids of Embryonic and Liver Zebrafish Cell Lines

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A 3D Culture Method of Spheroids of Embryonic and Liver Zebrafish Cell Lines

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January 20, 2023

DOI: 10.3791/64859-v

Irisdoris Rodrigues de Souza¹, Andrezza Di Pietro Micali Canavez², Desiree Cigaran Schuck², Viviana Stephanie Costa Gagosian¹, Isisdoris Rodrigues de Souza¹, Natália de Albuquerque Vita², Edvaldo da Silva Trindade³, Marta Margarete Cestari⁴, Marcio Lorencini², Daniela Morais Leme⁴

¹Graduate Program in Genetics, Department of Genetics,Federal University of Paraná (UFPR), ²Safety of Product Department,Grupo Boticário, ³Department of Cell Biology,Federal University of Paraná (UFPR), ⁴Department of Genetics,Federal University of Paraná (UFPR)

Overview

This article presents a simple and efficient protocol for generating 3D zebrafish spheroids using two cell lines: ZEM2S and ZFL. The method is designed for ease of use and rapid execution, making it suitable for various applications in fish toxicity testing.

Key Study Components

Area of Science

Neuroscience
Cell Biology
Toxicology

Background

Zebrafish are commonly used in biological research due to their transparent embryos and rapid development.
Spheroids serve as a 3D in vitro model that closely mimics in vivo conditions.
Understanding chemical effects on fish is crucial for environmental safety assessments.
This protocol aims to streamline the process of spheroid formation for research purposes.

Purpose of Study

To develop a fast and effective method for creating zebrafish spheroids.
To facilitate research on the effects of chemicals on fish.
To improve methodologies in fish toxicity testing.

Methods Used

Utilization of T75 flasks for cell culture.
Maintaining cells at approximately 80% confluence.
Following a step-by-step protocol for spheroid formation.
Demonstration by a genetics expert to ensure clarity.

Main Results

The protocol successfully generates uniform spheroids from zebrafish cell lines.
Spheroids exhibit consistent size and shape, suitable for experimental use.
Potential applications in assessing chemical toxicity in aquatic environments.
Streamlined process minimizes complications during spheroid formation.

Conclusions

This 3D culture protocol is effective for generating zebrafish spheroids.
It holds promise for enhancing fish toxicity testing methodologies.
Future research can leverage this model for environmental safety assessments.

Frequently Asked Questions

What are zebrafish spheroids used for?

Zebrafish spheroids are used as a 3D in vitro model for studying the effects of chemicals on fish and improving toxicity testing.

How long does it take to generate spheroids?

The protocol is designed to be fast, allowing for quick generation of spheroids when steps are followed correctly.

What cell lines are used in this protocol?

The protocol utilizes two zebrafish cell lines: ZEM2S (embryo) and ZFL (normal hepatocyte).

Who demonstrates the procedure?

The procedure is demonstrated by Irisdoris Rodrigues de Souza, a doctor in genetics.

What is the significance of using zebrafish in research?

Zebrafish are valuable in research due to their genetic similarity to humans and their transparent embryos, which facilitate observation.

Here, we present an effective, easy, and fast 3D culture protocol for the formation of spheroids of two zebrafish (Danio rerio) cell lines: ZEM2S (embryo) and ZFL (normal hepatocyte).

This protocol can easily generate zebrafish spheroids, a 3D in vitro model with different potential application in fish toxicity testing. This is a simple and fast method to generate single spheroids of zebrafish cell lines very similar in size and shape. If you follow all the described steps correctly, one should not encounter any problems.

Zebrafish spheroids provides may greatly benefit the mechanism, exploration of chemicals'effects on fish, which helps to improve the environmental safety assessment of products such as cosmetics. Demonstrating the procedure will be Irisdoris Rodrigues de Souza, a doctor in genetics from my laboratory. To begin, take a T75 flask of zebrafish cells at around 80%confluence.

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