Technologies and Appliances

 We generate high-titer third generation lentiviral vectors for in vitro and in vivo experiments. We routinely use these vectors for both overexpression and knock-down of genes using shRNA or CRISPR/Cas9 technology. Our viral vectors are generated by transient transfection of HEK293T producer cells with lentiviral constructs, followed by ultracentrifugation-based purification and concentration.

To study energy metabolism of adipocytes, we use numerous primary murine and human brown, beige and white cell models. To over express or knockdown genes of interest during the prolonged differentiation of these cells, we make use of high-titer lentiviral particles, which we produce in-house.

Investigating large libraries of small molecular compounds or silencing RNAs has been a staple in drug discovery for many years. In our lab, we use this methodology to uncover new pharmacology and/or metabolic pathways that could potentially aid us in the battle against obesity. By making use of state-of-the-art automation as, for example, machine-controlled precise cell seeding using the LLC FlowJo system and robotics-aided liquid handling using the Agilent Bravo platform, high-throughput examination of up to 10.000 substances becomes feasible. Accurate detection with our multimode absorbance-based PerkinElmer EnSpire and microscopy-based BioTek Cytation 5 plate-readers, allows analyses of a multifold of different assays and parameters.

The complete characterization of the metabolome requires a reliable identification of various metabolites in our samples, which is accurately addressed using high-resolution mass spectrometry. The metabolic products are quantitatively extracted from various biological sample matrices, separated chromatographically and sensitively determined using high-resolution mass spectrometry. So, individual metabolites can reliably be identified and quantified, and, moreover, complex metabolic cascades in the adipocytes can also be examined in detail.

The metabolism of higher organisms is regulated by concerted action of several organs like white adipose tissue, liver, skeletal muscle, or brown fat. To understand the role and impact of signaling cascades and if there is a therapeutic potential to increase energy expenditure - for example to treat obesity - we have to study whole organisms where interactions of different organs and/or tissues takes place. Therefore, we apply adipose tissue-specific mouse models and perform metabolic screening using metabolic cages of TSE Phenomaster Systems. These allow to measure metabolic parameters like O2 consumption or CO2 production at high temporal resolution and at defined temperatures over prolonged time.