Ruoß Marc, Kieber Vanessa, Rebholz Silas, Linnemann Caren, Rinderknecht Helen, Häussling Victor, Häcker Marina, Olde Damink Leon H H, Ehnert Sabrina, Nussler Andreas K
Department of Traumatology, Siegfried Weller Institute, BG-Klinik Tübingen, Eberhard Karls University, 72076 Tübingen, Germany.
Matricel GmbH, 52134 Herzogenrath, Germany.
Methods Protoc. 2019 Dec 23;3(1):1. doi: 10.3390/mps3010001.
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.
近年来，为了提高人肝细胞和肝癌细胞系的代谢活性，已有许多方法被报道。代谢活性的提高主要通过在三维系统或共培养（与其他细胞系）中培养细胞来实现。然而，如果系统变得更加复杂，对细胞数量进行量化就会变得更加困难（例如，在三维基质上）。到目前为止，在三维共培养系统中单独量化不同细胞类型是不可能的。因此，我们开发了一种基于聚合酶链式反应的方法，该方法能够在三维支架培养中分别对HepG2细胞和3T3-J2细胞进行量化。此外，我们的结果表明，与常用的基于代谢活性测量的方法（如刃天青的转化）相比，该方法能使二维和三维培养之间具有更好的可比性。
