Development of a stretch‐induced neurotrauma model for medium‐throughput screening in vitro: identification of rifampicin as a neuroprotectant

Abstract

This article is part of a themed section on Inventing New Therapies Without Reinventing the Wheel: The Power of Drug Repurposing. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v175.2/issuetoc.

In vitro modeling of dendritic atrophy in Rett syndrome: determinants for phenotypic drug screening in neurodevelopmental disorders

“…www.nature.com/scientificreports www.nature.com/scientificreports/ Previous in vitro mouse models reproducing the developmental deficits of the Rett syndrome were developed using high seeding cell-density in the range of 600-800 cells/mm 2 in 24-well plates 14,27,[30][31][32] . Although a variety of cell-seeding densities ranging from 640 to 40 cells/mm 2 are currently in use [33][34][35] , the available literature on phenotypic drug screening suggests that to perform a reliable measure of cellular morphology using automated image analysis tools, the cellular density should be low 18,36 . In neural cultures, low density ensures that neurites do not fasciculate extensively and then can be correctly identified and measured by an analysis software.…”

Proliferation and differentiation of human fetal brain neural stem cells in vitro

“…The ability to culture and manipulate hNSCs provides a critical tool that can be used for a variety of purposes from modeling human disease to high throughput drug screening. [6][7][10][11][12] One of the challenges with hNSC culture is maintaining the "stemness" of the culture. Svendsen et al developed a method in 1998 to maintain primary human fetal neural stem cells in culture by chopping neurospheres into pieces.…”

Cell-Based Screening to Identify Cytoprotective Compounds

“…Based on the assumptions that (1) the wells treated with various ineffective drugs will show similar response to the vehicle-treated wells and (2) there are very few active drugs, the compound-treated wells can quasi replace the negative control wells in the analysis. However, if the dataset shows larger variation, the Z-scores will be smaller values, thus meaningful cut-off values should be chosen individually for the selection of hits in each experimental model [54,58,59]. Apart from the more complex procedures, like Z-score calculation, there are various other options to identify cytoprotective compounds.…”

“…Either way, choosing the best model is more important than the use of human cells, since it is better to loose a few hits because of the differences in orthologs than to identify hits in an irrelevant model. If there are multiple options to choose from, it is worth keeping the lower cost model for the primary screening and use the other cell type as secondary model to confirm the action of hit compounds [58,59].…”

“…The dilution step of compounds may be associated with contamination risk, partly because the storage plates often use tube caps that require manual handling and cannot be removed by a robotic system or because the pipetting station used for the dilution step does not fit in a biosafety cabinet. Also, experimental devices that are necessary to induce cellular injury may be utilized in some models and these instruments may exceed the size limits of a laminar flow cabinet [59]. While contamination risk may be minimized by careful planning of experiments, it is often impossible to eliminate all sources of bacterial contaminants.…”