Single cell transcriptomics reveals reduced stress response in stem cells manipulated using localized electric fields

Membrane disruption using Bulk Electroporation (BEP) is a widely used non-viral method for delivering bio-molecules into cells. Recently, its microfluidic counterpart, Localized Electroporation (LEP), has been successfully used for several applications ranging from reprogramming and engineering cells for therapeutic purposes to non-destructive sampling from live cells for temporal analysis. However, the side effects of these processes on gene expression, that can affect the physiology of sensitive stem cells are not well understood. Here, we use single cell RNA sequencing (scRNA-seq) to investigate the effects of BEP and LEP on murine neural stem cell (NSC) gene expression. Our results indicate that unlike BEP, LEP does not lead to extensive cell death or activation of cell stress response pathways that may affect their long-term physiology. Additionally, our demonstrations show that LEP is suitable for multi-day delivery protocols as it enables better preservation of cell viability and integrity as compared to BEP.

Automated summary

We used single cell RNA sequencing to investigate the effects of BEP and LEP on gene expression in murine neural stem cells. Our results show that LEP does not lead to extensive cell death or activation of cell stress response pathways, making it suitable for long-term preservation of cell physiology. In addition, LEP enables better cell viability and integrity compared to BEP, making it a promising method for multi-day delivery protocols.