Q3 Prof. Dr. Weiskirchen
FACS-based cell isolation and clinical translation

The Q3 Core Unit provides the projects with reliably, standardized and efficiently isolated primary cells from liver and kidney from mice and rats. We have established FACS-based protocols for isolating hepatic stellate cells, sinusoidal endothelium as well as renal podocytes and parietal cells. We are currently extending our protocols to human cell types for translational projects.

This core facility established innovative procedures that provide researchers of the SFB/TRR57 with healthy viable primary cells of murine and rat livers and kidneys in a very reliable, standardised and cost-effective centralised manner. Using conventional isolation procedures, we have set up a logistical and methodological platform that routinely provides primary hepatic stellate cells and hepatocytes to all projects in Aachen and Bonn upon request. Moreover, we have introduced novel FACS-based sorting protocols that allow isolation of highly purified hepatic stellate cells from murine livers using their autofluorescence in the UV channel and negativity for hematopoietic surface markers. Other protocols, both FACS- and MACS-based, were established for the isolation of different kidney cell types (podocytes, parietal cells, macrophages/dendritic cells), hepatic sinusoidal endothelial cells, oval cells as well as immune cells from liver and kidney from mice and rats.

We now expand our methodology to additionally provide the infrastructure for working with primary human liver cells to facilitate translational research within the SFB/TRR 57. Dr. Jeanette Bierwolf (Dept. of Visceral Surgery, UKB) will provide human hepatocytes from liver resections for individual projects upon request. Furthermore, we have started to establish techniques to isolate immune cell populations (macrophage subsets, lymphocytes) from human liver, based on the BMBF-funded biobanking program at the University Hospital Aachen.

We have furthermore published methodological papers on hepatic endothelial and stellate cell isolation techniques, thereby providing peer-reviewed references for the individual projects when referring to the methodology. During the last years, we have organized several internal methodological training courses for students and researchers within the SFB/TRR57 and hosted several national and international external scientists that were trained in these novel isolation techniques.