Methodology

Genetic methods:

  • Genotyping using PCR methodology as well as real-time PCR techniques (e.g. TaqMan assays).
  • Gene expression profiling (blood, tissue samples, postmortem, animal models, cell culture, iPSC etc.) using quantitative real-time RT-PCR methodology and microarrays.
  • In situ hybridization methodology for localization.
  • Manipulation of gene expression in vitro using siRNA as well as plasmids.

Biochemical analysis:

  • Protein analysis via western blot, ELISA and immunohistochemical/immunocytochemical analysis.
  • Fluorescence and colorimetric cytochemical analysis and evaluation via computerized software for density and localization (microscopically).
  • Enzymatic activity assays – influence of therapy and genes.

Cell culture models:

  • Neuronal cell lines treatments for differentiation, proliferation and cell death.
  • Primary cell culture for drug therapy study.
  • Lymphoblastoid cell culture originating from cases and controls for molecular mechanisms investigations.
  • Cell line models using siRNA / plasmid methodology for down/up regulation of a specific gene.
  • Live Cell imaging techniques for monitoring changes in neuronal cell culture.

Induced Pluripotent stem cells (iPSC):

  • Extractions and cultivation of keratinocytes from patients and health controls from hair follicles.
  • Reprogramming of keratinocytes into iPSC followed by differentiation into neuronal progenitor cells and matured neuronal cells.
  • Investigation of molecular mechanisms and therapy treatment effects on the patients based neuronal cells originating from the iPSC.

Electrophysiology

Cell Analyzer Tool:

In the following movie a recent method is introduced, developed and evaluated in our lab. You can also download the associated paper below.

Real-Time Impedance-based Cell Analyzer as a Tool to Delineate Molecular Pathways Involved in Neurotoxicity and Neuroprotection in a Neuronal Cell Line

Zoya Marinova, Susanne Walitza, Edna Grünblatt