Genetic methods:
- Genotyping using PCR methodology, real-time PCR techniques (e.g. TaqMan assays) and Genome-wide association studies (GWAS) with subsequent polygenic risk scores (PRS) analysis.
- Gene expression profiling (blood, tissue samples, postmortem, animal models, cell culture, iPSC etc.) using quantitative real-time RT-PCR methodology, microarrays and RNA sequencing (in collaboration).
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.
- 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 cultures from both cases and controls are used to study molecular mechanisms.
- Cell line models using siRNA / plasmid methodology for down/up regulation of a specific gene.
- Reporter assays.
- Live Cell imaging techniques for monitoring changes in neuronal cell culture.
Induced Pluripotent stem cells (iPSC):
- Isolating and growing keratinocytes and PBMCs from hair follicles and blood samples obtained from patients and healthy individuals.
- Somatic cells reprogrammed into iPSCs, then differentiated into neural progenitor cells and mature neuronal cells.
- Studying the molecular causes and therapeutic impacts on patients using neural cells derived from iPSCs.
Electrophysiology:
• In-vitro neural activity and synchronised network recordings are preformed using a multichannel system (MEA2100 and Multiwell-ME).
• Collaborative HD-MEA measurements are carried out with Prof. Andreas Hierlemann (ETHZ).
Cell Analyzer Tool:
The xCELLigence real-time impedance cell analyzer is utilised to monitor adherent cell growth or death continuously for hours and days under different treatments or manipulations.
In the subsequent paper, the method was introduced, developed, and assessed in our laboratory. You may also download the accompanying document 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 Marinova, Z., Walitza, S., & Grünblatt, E. (2014).
Real-Time Impedance-based Cell Analyzer as a Tool to Delineate Molecular Pathways Involved in Neurotoxicity and Neuroprotection in a Neuronal Cell Line. JoVE, (90) (PDF, 966 KB)
