Single Cell Biology

Abstract

On the Generation of Efficient Methodologies for Cell-Based Toxicity Screening by Comparative Protocol Analysis and Optimization

Sepideh Abolpour Mofrad, Katharina Kuenzel, Oliver Friedrich and Daniel F Gilbert*

In cell biology in general and in high-throughput cell-based screening approaches in particular - e.g. for in vitrobased toxicity assessment - various methodologies or protocols are reported in the literature. In the case that a cell-based screening assay, for toxicity evaluation or for assessing another biological question, is to be established for the first time in a research lab, the researcher has to select a number of different protocols from the literature and to create an optimized protocol for the intended use. This is typically required, because optimized protocols, generated following comparative protocol analysis and optimization, are mostly rarely available. In another study, which we refer to as a showcase, we conducted a comparative analysis of three different protocols for neuronal NT2 cell differentiation, available in the literature. From this comparison, we generated an improved and optimized method, allowing for neuronal NT2 differentiation in monolayer cultures with high yield of NT2-N cells, allowing for systematic in vitro-based primary screening for developmental toxicants and neuro-toxicants at different stages of maturation. In this commentary, to prevent the same experiments from being repeatedly conducted in different labs around the world and at different times over and over again, we suggest generation of advanced and efficient methods by comparative protocol analysis and optimization, for application in a variety of research fields, related to cell and single cell biology.