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

• Main 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
• Format: Article
• Language: English
• Published: MDPI AG 2019-12-01
• Series: Methods and Protocols
• Subjects: quantification; 3d culture; co-culture; cell number; pcr-based method
• Online Access: https://www.mdpi.com/2409-9279/3/1/1

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.