Eurofins Discovery provides phenotypic cell-based functional assays to screen agents for wound healing, neurite outgrowth, and angiogenesis applications. Phenotypic cell-based drug discovery approaches model a specific biology with physiological and pathological relevance. There is currently a resurgence in phenotypic drug discovery based on advanced cellular assays and their potential to reveal polypharmacology, address complex biology, and deliver first-in-class medicines.
Neurodegenerative disorders are a challenging therapeutic area and an unmet medical need, especially for prevalent diseases such as Alzheimer’s disease (AD) and Parkinson’s disease (PD). Genetic variants identified as contributors to disease progression and neurodegeneration include β-amyloid/ hyperphosphorylated-tau in familial AD, APOE4 in sporadic AD, and α-synuclein in PD progression. To combat these diseases, our scientists have developed human neurite outgrowth models based on human neuroblastoma SH-SY5Y cells carrying these risk gene variants. Relevant assays are available individually or as part of a comprehensive Neurodegeneration Phenotypic Panel.
Figure 1. Representative neurite morphology of SH-SY5Y genetic variant cells. A. Fluorescent image of SY-SY5Y cells expressing APP-PS1-MAPT co-expressed with red fluorescent protein (RFP) for quantitative analysis on neurite outgrowth. Morphology is shown by RFP (red) and nuclei (blue) counterstained with Hoechst 33342 (10X objective). B. Dose response curve of staurosporine treatment is shown for neurite outgrowth measurement.
| Assay | Genetic variant | Clinical relevance | Item # |
|---|---|---|---|
| α-Synuclein A53T, Neurite Outgrowth | α-SynucleinA53T | PD | 332400-0 |
| APOEε4 C130R, Neurite Outgrowth | APOEε4C130R | Sporadic AD, late onset | 332440-0 |
| APP-PS1-MAPT, Neurite Outgrowth | APPSwe/Lon-PS1ΔE9-MAPT 2N4R-P301L | Familial AD, early onset | 332460-0 |
- Our Neurodegeneration Phenotypic Panel (Item PP283) provides an in vitro, cell-based exploration of neurite outgrowth in a model carrying disease-relevant high risk genes
- Integrated, customized image analysis modules enable unbiased analysis of complex phenotypic changes
- Robust assays deliver easy scale-up and reproducible data
Keratinocyte-based Wound Healing Assay
Our cell migration assays solve the challenges of inconsistency, primary cell donor variability, and lack of automated quantitation. Our assays uses immortalized human keratinocytes (HaCaT cells) in a cell exclusion zone rather than in a scratch assay, to provide reliable and reproducible results. While primary human keratinocytes are highly biologically relevant, their relatively short culture lifetime and donor to donor variability create a challenge for data reproducibiity. For unbiased quantitative analysis, our assays incorporate confocal High Content Analysis to measure the decrease of closure area, with test agent induced cell migration calculated relative to an EGF control response.
Figure 2. Keratinocytes and fibroblasts play an important role in the proliferative phase of wound healing, to restore tissue structure and function. Activation of keratinocytes and fibroblasts by macrophages, cytokines, and growth factors cause angiogenesis, extracellular matrix changes, and epithelialization.
Angiogenesis Tube Formation Assay for Biological Relevance
Angiogenesis (blood vessel formation) is essential in tissue growth, development, and the progression of many human malignancies and life-threatening diseases. Anti-angiogenic therapy is a novel and effective approach for treatment of angiogenesis-dependent diseases such as cancer, diabetic retinopathy, and age-related macular degeneration. Our Eurofins Discovery experts developed a tube formation model that utilizes human umbilical vein endothelial cells (HUVECs) self-organized in a 3D network structure in matrigel. Our network structure model provides richer information with higher translational relevance compared to target-based and 2D cellular models.
Figure 3. Data-rich high content imaging provides maximum value from each assay well. Traditional tube formation model (left) with limited view and missing information. Eurofins Discovery tube formation model (right) utilizing fluorescence-labeled HUVEC cells imaged by confocal microscopy and analyzed with Z-stack projection. High content assays provide whole-well field of view imaging and analysis for maximum informational detail.
