Progressing 3D Spheroid Analysis into a HTS Drug Discovery Method
Poster Dec 04, 2014
Sarah Kessel, Eric Sincoff, Olivier Dery
Inhibition of cancer cell proliferation in drug discovery research has not translated well from traditional two-dimensional (2D) in vitro assays to in vivo studies. Most anti-cancer drug compound studies are performed in a tissue culture treated 2D assay format for the purpose of studying proliferation, viability, and apoptosis. Increasingly, scientific evidence is showing that growing cancer cells in the form of three-dimensional (3D) spheroids is more predictive of in vivo study outcomes than 2D cell culture formats. Typically, spheroids are created using various methods, such as U-bottom plates, hanging drop or semi solid media and the analysis of sphere growth has been carried out using a microscope, which is time consuming, laborious, low-throughput and lacks reproducibility. Literature has previously reported the use of an automated imager to monitor the size and growth of 3D spheroids in 96-well plates (1), where spheres were formed in 4 days from a set number of cells to return a consistent sphere size. This study highlights the use of the 384-well low attachment U-bottom plate combined with the Celigo® Imaging Cytometer to image and analyze the formation of 3D spheroids. This allows for increased throughput, number of replicates and parameters per plate as compared to 96-well plates. U87MG cells were used to create tumorspheres in 384-well plates and were subsequently analyzed by imaging. The data illustrate that reproducible 3D spheroids can be formed in 384-well plates. Fluorescent viability studies were carried out with the imager using pixel intensity analysis. Moreover, the assay was validated for drug screen using various drug compounds that have shown antiproliferative effects. Together, these data demonstrate that the tumorsphere formation assay can be developed, validated and used for high-throughput anticancer compound screening in 384-well U-bottom low attachment plates using the Celigo Imaging Cytometer.
Extracorporeal shockwave therapy accelerates motor axon regeneration despite a phenotypically mismatched environmentPoster
A femoral nerve defect model was adapted for the evaluation of proregenerative effects of extracorporeal shockwave therapy (ESWT). Functional evaluation, histology and qRT-PCR data show differences between sensory and motor-derived nerve transplants and a pro-regenerative effect of ESWT. These data provide evidence for the clinical application of ESWT after autologous nerve transplantation as a novel non-invasive method.READ MORE
Cerebral Malaria Insights: Pathogenesis, Host Parasite Interactions including Host ResistancePoster
Cerebral malaria is a dreadful disease transmitted by mosquito. The major preventive approach is focused more in vector control than development of anti-malarial drug. The purpose of this presentation is to analyze different aspects of disease manifestations including clinical symptoms and pathogenesis in the context of mosquito borne infections in different geographical regions of the world.READ MORE