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Cell-Type-Specific Quantification of a Scaffold-Based 3D Liver Co-Culture

Authors :
Marc Ruoß
Vanessa Kieber
Silas Rebholz
Caren Linnemann
Helen Rinderknecht
Victor Häussling
Marina Häcker
Leon H. H. Olde Damink
Sabrina Ehnert
Andreas K. Nussler
Source :
Methods and Protocols, Vol 3, Iss 1, p 1 (2019)
Publication Year :
2019
Publisher :
MDPI AG, 2019.

Abstract

In order to increase the metabolic activity of human hepatocytes and liver cancer cell lines, many approaches have been reported in recent years. The metabolic activity could be increased mainly by cultivating the cells in 3D systems or co-cultures (with other cell lines). However, if the system becomes more complex, it gets more difficult to quantify the number of cells (e.g., on a 3D matrix). Until now, it has been impossible to quantify different cell types individually in 3D co-culture systems. Therefore, we developed a PCR-based method that allows the quantification of HepG2 cells and 3T3-J2 cells separately in a 3D scaffold culture. Moreover, our results show that this method allows better comparability between 2D and 3D cultures in comparison to the often-used approaches based on metabolic activity measurements, such as the conversion of resazurin.

Details

Language :
English
ISSN :
24099279
Volume :
3
Issue :
1
Database :
Directory of Open Access Journals
Journal :
Methods and Protocols
Publication Type :
Academic Journal
Accession number :
edsdoj.f8a0e02bc5174a4fb5a845048f76f8eb
Document Type :
article
Full Text :
https://doi.org/10.3390/mps3010001