Development of a novel high-throughput culture system for hypoxic 3D hydrogel cell culture

authored by: Dominik Egger, Luisa Baier, Julia Moldaschl, Manfred Taschner, Volker Lorber, Cornelia Kasper
Abstract: Animal models lack physiologic relevance to the human system which results in low clinical translation of results derived from animal testing. Besides spheroids or organoids, hydrogel-based 3D in vitro models are used to mimic the in vivo situation increasing the relevance while reducing animal testing. However, to establish hydrogel-based 3D models in applications such as drug development or personalized medicine, high-throughput culture systems are required. Furthermore, the integration of oxygen-reduced (hypoxic) conditions has become increasingly important to establish more physiologic culture models. Therefore, we developed a platform technology for the high-throughput generation of miniaturized hydrogels for 3D cell culture. The Oli-Up system is based on the shape of a well-plate and allows for the parallel culture of 48 hydrogel samples, each with a volume of 15 µl. As a proof-of-concept, we established a 3D culture of gelatin-methacryloyl (GelMA)-encapsulated mesenchymal stem/stromal cells (MSCs). We used a hypoxia reporter cell line to establish a defined oxygen-reduced environment to precisely trigger cellular responses characteristic of hypoxia in MSCs. In detail, the expression of hypoxia response element (HRE) increased dependent on the oxygen concentration and cell density. Furthermore, MSCs displayed an altered glucose metabolism and increased VEGF secretion upon oxygen-reduction. In conclusion, the Oli-Up system is a platform technology for the high-throughput culture of hydrogel-based 3D models in a defined oxygen environment. As it is amenable for automation, it holds the potential for high-throughput screening applications such as drug development and testing in more physiologic 3D in vitro tissue models.
Organisation(s): Institute of Cell Biology and Biophysics
Biofabrication for drug testing
External Organisation(s): University of Natural Resources and Applied Life Sciences (BOKU)
LifeTaq-Analytics GmbH
Type: Article
Journal: Scientific reports
Volume: 14
No. of pages: 9
ISSN: 2045-2322
Publication date: 30.04.2024
Publication status: Published
Peer reviewed: Yes
ASJC Scopus subject areas: General
Sustainable Development Goals: SDG 3 - Good Health and Well-being
Electronic version(s): https://doi.org/10.1038/s41598-024-60822-z (Access: Open)

BibTeX

@article{1ebf82e99c7045eab61b0dc6c8cf1f11,
title = "Development of a novel high-throughput culture system for hypoxic 3D hydrogel cell culture",
abstract = "Animal models lack physiologic relevance to the human system which results in low clinical translation of results derived from animal testing. Besides spheroids or organoids, hydrogel-based 3D in vitro models are used to mimic the in vivo situation increasing the relevance while reducing animal testing. However, to establish hydrogel-based 3D models in applications such as drug development or personalized medicine, high-throughput culture systems are required. Furthermore, the integration of oxygen-reduced (hypoxic) conditions has become increasingly important to establish more physiologic culture models. Therefore, we developed a platform technology for the high-throughput generation of miniaturized hydrogels for 3D cell culture. The Oli-Up system is based on the shape of a well-plate and allows for the parallel culture of 48 hydrogel samples, each with a volume of 15 µl. As a proof-of-concept, we established a 3D culture of gelatin-methacryloyl (GelMA)-encapsulated mesenchymal stem/stromal cells (MSCs). We used a hypoxia reporter cell line to establish a defined oxygen-reduced environment to precisely trigger cellular responses characteristic of hypoxia in MSCs. In detail, the expression of hypoxia response element (HRE) increased dependent on the oxygen concentration and cell density. Furthermore, MSCs displayed an altered glucose metabolism and increased VEGF secretion upon oxygen-reduction. In conclusion, the Oli-Up system is a platform technology for the high-throughput culture of hydrogel-based 3D models in a defined oxygen environment. As it is amenable for automation, it holds the potential for high-throughput screening applications such as drug development and testing in more physiologic 3D in vitro tissue models.",
author = "Dominik Egger and Luisa Baier and Julia Moldaschl and Manfred Taschner and Volker Lorber and Cornelia Kasper",
note = "Funding Information: Open Access funding enabled and organized by Projekt DEAL. This study was supported by the Austrian Research Promotion Agency (FFG) grant number 880720. ",
year = "2024",
month = apr,
day = "30",
doi = "10.1038/s41598-024-60822-z",
language = "English",
volume = "14",
journal = "Scientific reports",
issn = "2045-2322",
publisher = "Nature Publishing Group",
number = "1",
}