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Um inserto inovador impresso em 3D projetado para permitir culturas de células 2D e 3D diretas
Um inserto inovador impresso em 3D projetado para permitir culturas de células 2D e 3D diretas
JoVE Journal
Biology
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JoVE Journal Biology
An Innovative 3D-Printed Insert Designed to Enable Straightforward 2D and 3D Cell Cultures

Um inserto inovador impresso em 3D projetado para permitir culturas de células 2D e 3D diretas

Full Text
1,906 Views
08:17 min
January 6, 2023

DOI: 10.3791/64655-v

Charlie Colin-Pierre1,2,3, Oussama El Baraka3, Laurent Ramont1,2,4, Stéphane Brézillon1,2

1Laboratoire de Biochimie Médicale et Biologie Moléculaire,Université de Reims Champagne-Ardenne, 2UMR CNRS 7369, Matrice Extracellulaire et Dynamique Cellulaire-MEDyC,Université de Reims Champagne-Ardenne Reims, 3BASF Beauty Care Solutions, 4Service Biochimie-Pharmacologie-Toxicologie,Centre Hospitalier Universitaire (CHU) de Reims

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Overview

This study presents a novel 3D-printed insert model to investigate co-culture systems, focusing on the paracrine intercellular communication between endothelial cells and keratinocytes. The insert enhances experimental model design for various cell types and demonstrates significant increases in endothelial cell proliferation and migration when co-cultured with keratinocytes.

Key Study Components

Research Area

  • Cell biology
  • Endothelial cell-keratinocyte interactions
  • 3D culture models

Background

  • The importance of direct cell communication in biological systems.
  • Challenges in establishing culture models for different cell types.
  • Need for scalable, flexible cell culture techniques.

Methods Used

  • Use of a 3D-printed insert for cell culture.
  • Co-culture of keratinocytes and human dermal microvascular endothelial cells.
  • Analysis of cell viability and communication effects through migration and proliferation assays.

Main Results

  • Keratinocytes significantly enhanced endothelial cell proliferation by 1.5-fold after 24 hours and by 3.1-fold after 48 hours.
  • Endothelial cell migration improved significantly in the presence of keratinocytes, covering up to 99% of the wound area by 24 hours.
  • High cell viability was observed in the 3D-printed inserts compared to conventional culture methods.

Conclusions

  • This study demonstrates the efficacy of 3D-printed inserts in facilitating direct cell interactions and culture.
  • The findings have implications for understanding cell communication and can aid in studying various physiopathologies.

Frequently Asked Questions

What are the advantages of using 3D-printed inserts in cell culture?
3D-printed inserts are durable, flexible, and scalable, allowing for improved experimental models for various studies.
How do keratinocytes affect endothelial cell behavior?
Keratinocytes significantly enhance endothelial cell proliferation and migration, demonstrating important intercellular communication.
What is the main application of this co-culture model?
It is applicable to studying direct cell communication and various physiopathologies including immunology.
What are the key findings related to cell viability?
Cell viability rates were high, with 90% in 6-well plates and 91% in the 3D-printed inserts for keratinocytes.
Can this model be used for other cell types?
Yes, the 3D-printed inserts can be modified for various cell types requiring different coatings.
What protocols are followed for preparing the 3D inserts?
The inserts are created by mixing silicone and catalyst, sterilized, and then incubated for cell attachment and growth.

Neste artigo, um novo inserto impresso em 3D é apresentado como um modelo de co-cultura e validado através do estudo da comunicação intercelular parácrina entre células endoteliais e queratinócitos.

Este protocolo pode ser usado para investigar numerosos estudos de cultura e fornecer uma nova ferramenta para investigar a comunicação celular direta. Este novo dispositivo economiza tempo significativo no estabelecimento de um modelo de cultura e é aplicável a diferentes tipos de células que requerem um revestimento específico ou não. De fato, os quatro compartimentos das inserções impressas em 3D permitem cultivar diferentes tipos de células em monocamada, ou em 3D da mesma forma com diferentes combinações.

As pastilhas impressas em 3D são mais duráveis, flexíveis, escaláveis e podem ser usadas para projetar modelos experimentais para estudos de fisiopatologias, como imunologia ou androgênese. Para começar, misture os dois componentes, silício alimentar e catalisador fornecidos no kit na proporção de 10:1, usando uma espátula esterilizada a etanol 70% de acordo com as instruções do fabricante. Aplique as pastilhas sobre a mistura de silicone com uma pinça para que a mistura seja distribuída homogeneamente na borda inferior da pastilha.

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