In-vitro Pharmacology

A wide variety of in vitro tests are available within the Fraunhofer IME Project Group Translational Medicine and Pharmacology. Emphasis is laid on cellular signaling processes, lipid metabolism, neuronal cell responses and immune cell activity. Cytotoxicity, proliferation and apoptosis assays are performed. State-of-the art cellular, immunological, biochemical, analytical, proteomic and genomic techniques are employed.

TECHNIQUES AND METHODS

Cell culture techniques

  • Generation of plasmids and lenti virus expressing specific target genes
  • Transient and stable transfection/transduction of plasmids/lenti virus
  • Apoptosis assay, proliferation assay, migration assay, adhesion assay

Imaging methods

  • Imaging of proteins and mRNA in murine tissue sections (e.g. spinal cord, brain, lymph nodes) by in-situ hybridization and immunohistochemistry
  • Imaging of up to 15 proteins on one tissue section (Multi-Epitope-Ligand-Cartography)
  • Imaging of proteins in human and murine cells by flow cytometry

Analysis of lipid metabolism

  • LC-MS/MS of lipid mediators such as sphingolipids, eicosanoids, endocannabinoids, lysophosphatidic acids.

 

TEST SYSTEMS AND ASSAYS

Immune cell function test systems

  • Differentiation/activation of human dendritic cells
  • Differentiation/polarization of human M1-/M2-Macrophages
  • Activation/proliferation of human T-cells
    • Readouts: Change in morphology, Cytokine-/Chemokine (mRNA/protein level) determination, surface marker determination (flow cytometry), inflammatory mediator analysis (e.g. NO, PGE2)

Activity/Signaling assays

  • Characterization of Ca2+-dependent channels and receptors (calcium imaging)
  • Activity assays of proteins (e.g. NOX4-assay, BLT2-assay)
  • Receptor-binding assays (e.g. G-protein linked, eicosanoid receptors)
  • Promotor-Reporter-Gene assays
  • Development of customer specific assays

Selected Publications

Barthelmes J, Männer de Bazo M, Pewzner-Jung Y, Schmitz K, Mayer CA, Foerch C, Eberle M, Tafferner N, Ferreirós N, Birod K, Geisslinger G, Futerman A H, Grösch S and Schiffmann S (2015) Lack of CerS2 suppresses the development of experimental autoimmune encephalomyelitis by impairing the migratory capacity of neutrophils. Brain Behav Immun 46:280-92. doi: 10.1016/j.bbi.2015.02.010.

Knape T, Flesch D, Kuchler L, Sha LK, Giegerich AK, Labocha S, Ferreirós N, Schmid T, Wurglics M, Schubert-Zsilavecz M, Proschak E, Brüne B, Parnham MJ, von Knethen A (2015) Identification and characterisation of a prototype for a new class of competitive PPARγ antagonists. Eur J Pharmacol 755:16-26.

Kallenborn-Gerhardt W, Schröder K, Geisslinger G, Schmidtko A (2013) NOXious signaling in pain processing. Pharmacol Ther 137:309-17.