Neuronal development, function and the subsequent degeneration of the brain are still an enigma in both the normal and pathologic states and there is an urgent need to find better targets for developing therapeutic intervention. Current techniques to deconstruct the architecture of brain and disease related pathways are best suited for following up on single genes, but would take an impractical amount of time for the leads from the current wave of genetic and genomic data.
The aim of the cellomics facility is to provide a state of the art screening platform that conducts high-throughput and high-content (HT-HC) cellular screens. In combination with a whole genome human shRNA library one is able to perturb the function of hundreds of genes to identify enhancers and suppressors of phenotypes related to various neurodegenerative diseases, such as Parkinson's Disease, Alzheimer's Disease and Frontotemporal dementia.
This approach can contextualize the information, gathered from a combination of genetic and genomic approaches, into networks and functional biology and can be utilized for the identification of therapeutic targets.
Key aspects of the automated cell culture system:
To detect the effects of important genes/modifiers, it is essential to decrease experimental variation while minimizing error and costs. Our system allows the hands free simultaneous culturing of several mammalian cell lines, as key parameters (e.g. cell density, trypsinization times, confluency) can be adjusted independently for each cell type. This flexibility and throughput allows one to run several HT-HC experiments in parallel.
The above system has several pre-designed protocols to facilitate HT-HC screens in multiple cellular models:
- Adherent cell expansion and assay plate preparation
- Automated virus production, storage and delivery into the assay plates
- Automated expansion of mouse embyronic fibroblasts to faciliate iPS culture
- Expansion and assay plate preparation of embryonic stem cells
- Input and preparation of assay plates with more difficult cell lines (e.g. primary neurons, hiPS)
- Automated cellular differentiation.
- Fully programmable for the developmemt of new protocols.
In addition, the integration of two high content imagers (Operetta; Perkin Elmer, CV7000; Yokogawa) also allows for the quantification of numerous cellular phenotypes important for neurodegenerative diseases (Figure 1).
DZNE members, collaboration partners and external research groups or companies can apply for a screening or automation projects. Please contact Dr. Joachim Täger to arrange a meeting to discuss projects, use of equipment and pricing structure.