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

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

MethodsX (2020-10-03)
Jeffrey Aalders, Laurens Léger, Tim Tuerlings, Sergio Ledda, Jolanda van Hengel
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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Albumin human, Cellastim S, recombinant, expressed in rice, lyophilized powder, suitable for cell culture, low endotoxin, ≥96% (SDS-PAGE)
Sigma-Aldrich
L-Ascorbic acid 2-phosphate sesquimagnesium salt hydrate, ≥95%
Sigma-Aldrich
Wnt Antagonist II, IWP-2, The Wnt Antagonist II, IWP-2, also referenced under CAS 686770-61-6, controls the biological activity of Wnt. This small molecule/inhibitor is primarily used for Cancer applications.
Sigma-Aldrich
GSK-3 Inhibitor XVI, GSK-3 Inhibitor XVI - CAS 252917-06-9, is a cell-permeable, potent, ATP-competitive, and highly selective GSK-3 inhibitor (IC₅₀ = 10 and 6.7 nM against GSK-3α and GSK-3β, respectively).