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El Jalkh, T.

Publications and source records attributed to El Jalkh, T..

2 recordsLinked to original sources

Reversible and reusable compartmentalized microfluidic chip for coculture of dorsal root ganglion neurons

Compartmentalized microfluidic chips play an important role in understanding the cellular mechanisms involved in neurodegenerative disorders. Dorsal root ganglia are a well-established model for modelling the peripheral nervous system (PNS), but their development on a chip remains limited. Furthermore, it would be beneficial for the devices to be openable in order to access the biological material inside for analyses. Easy to prototype and biocompatible, styrenic block copolymers (SBC) are an alternative to polydimethylsiloxane (PDMS) that offer both reversible and permanent bonding properties. This paper presents a fast and straightforward method to produce compartmentalized SBC chips. The study validates the culture of murine dorsal root ganglia explants, comparing it to the standard methods, to obtain a model of the PNS. Moreover, the reversible bonding properties of the SBC permit the reuse of the chip with a quick and easy cleaning protocol. It provides direct access to the cells, opening the way for imaging and molecular biology analysis. The comparison of the resources required to produce PDMS and SBC chips highlights the importance of moving to reusable devices. These detachable, easy-to-manufacture and sustainable all-thermoplastic platforms provide an alternative way of prototyping compartmentalized devices for in vitro PNS modeling.

biophysics↗

RNA-Seq is not required to determine stable reference genes for qPCR normalization

Assessment of differential gene expression by qPCR is heavily influenced by the choice of reference genes. Although numerous statistical approaches have been proposed to determine the best reference genes, they can give rise to conflicting results depending on experimental conditions. Hence, recent studies propose the use of RNA-Seq to identify stable genes followed by the application of different statistical approaches to determine the best set of reference genes for qPCR data normalization. In this study, we demonstrate that the statistical approach to determine the best reference genes from randomly selected candidates is more important than the preselection of stable candidates from RNA-Seq data. Using a qPCR data normalization workflow that we have previously established; we show that qPCR data normalization using randomly chosen conventional reference genes renders the same results as stable reference genes selected from RNA-Seq data. We validated these observations in two distinct cross-sectional experimental conditions involving human iPSC derived microglial cells and mouse sciatic nerves. These results taken together show that given a robust statistical approach for reference gene selection, stable genes selected from RNA-Seq data do not offer any significant advantage over commonly used reference genes for normalizing qPCR assays.

molecular biology↗