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Biology
创新的 3D 打印插入物,旨在实现简单的 2D 和 3D 细胞培养
创新的 3D 打印插入物,旨在实现简单的 2D 和 3D 细胞培养
JoVE Journal
Biology
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JoVE Journal Biology
An Innovative 3D-Printed Insert Designed to Enable Straightforward 2D and 3D Cell Cultures

创新的 3D 打印插入物,旨在实现简单的 2D 和 3D 细胞培养

Full Text
1,901 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.

在本文中,提出了一种新设计的3D打印插入物作为共培养模型,并通过研究内皮细胞和角质形成细胞之间的旁分泌细胞间通讯进行了验证。

该协议可用于研究许多培养研究,并为研究直接细胞通讯提供新工具。这种新设备节省了建立培养模型的大量时间,并且适用于是否需要特定涂层的不同细胞类型。事实上,3D打印插入物的四个隔室允许以单层或以相同的方式以不同的组合培养不同的细胞类型。

3D打印的插入物更耐用,灵活,可扩展,可用于设计生理病理学研究的实验模型,例如免疫学或雄激素生成。首先,根据制造商的说明,使用70%乙醇灭菌刮刀,以10:1的比例混合试剂盒中提供的两种成分,食品硅和催化剂。用镊子将嵌件涂在硅胶混合物上,使混合物均匀分布在嵌件的底部边缘。

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