Liquid marble technology to create cost-effective 3D cardiospheres as a platform for in vitro drug testing and disease modelling

Jeffrey Aalders,Laurens Léger,Tim Tuerlings,Sergio Ledda,Jolanda Van Hengel

doi:10.1016/j.mex.2020.101065

Jeffrey Aalders, Laurens Léger + Show 3 more

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https://doi.org/10.1016/j.mex.2020.101065

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Journal: MethodsX	Publication Date: Jan 1, 2020
Citations: 9	License type: cc-by

Affiliation: Ghent University, University of Sassari

Abstract

Three-dimensional (3D) cell culturing has several advantages over 2D cultures. 3D cell cultures more accurately mimic the in vivo environment, which is vital to obtain reliable results in disease modelling and toxicity testing. With the introduction of the Yamanaka factors, reprogramming of somatic cells to induced pluripotent stem cells (iPSCs) became available. This iPSC technology provides a scalable source of differentiated cells. iPSCs can be programmed to differentiate into any cell type of the body, including cardiomyocytes. These heart-specific muscle cells, can then serve as a model for therapeutic drug screening or assay development. Current methods to achieve multicellular spheroids by 3D cell cultures, such as hanging drop and spinner flasks are expensive, time-consuming and require specialized materials and training. Hydrophobic powders can be used to create a micro environment for cell cultures, which are termed liquid marbles (LM). In this procedure we describe the first use of the LM technology for 3D culturing in vitro derived human cardiomyocytes which results in the formation of cardiospheres within 24h. The cardiospheres could be used for several in depth and high-throughput analyses.

Highlights

Method ArticleJeffrey Aalders a, Laurens Léger a, Tim Tuerlings a, Sergio Ledda b, Jolanda van Hengel a,∗
Immunofluorescent staining revealed that cardiac markers were retained after 3D culture in liquid marbles (LM) for all conditions (Fig. 2C) similar as cardiomyocytes cultured in 2D (Fig. 2B)
Depending on the experimental needs and the properties of the cell line, optimal cell suspension concentrations may vary. In this protocol we describe the LM technique with the broadly used H9 line that in our hands works comparable for human induced pluripotent stem cells

Summary

Method Article

Jeffrey Aalders a, Laurens Léger a, Tim Tuerlings a, Sergio Ledda b, Jolanda van Hengel a,∗. Hydrophobic powders can be used to create a micro environment for cell cultures, which are termed liquid marbles (LM) In this procedure we describe the first use of the LM technology for 3D culturing in vitro derived human cardiomyocytes which results in the formation of cardiospheres within 24h. The majority of the 3D technologies require specialized and expensive equipment, extensive experience, are time-consuming and have a relatively low reproducibility In this protocol we describe a robust and cost-effective method for the generation of 3D in vitro cardiac cultures that does not require specialized equipment. The use of hydrophobic powders in LM was first described by Aussillous and Quéré in 2001 [7] This technology provides a method to create a micro bioreactor for 3D cell culture. Applications of the LM have extended to the broader field of biology including the use in enhancing polymerase chain reactions [13]

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