TY - JOUR
T1 - Single cell transcriptomics reveals reduced stress response in stem cells manipulated using localized electric fields
AU - Mukherjee, Prithvijit
AU - Peng, Chian Yu
AU - McGuire, Tammy
AU - Hwang, Jin Wook
AU - Puritz, Connor H.
AU - Pathak, Nibir
AU - Patino, Cesar A.
AU - Braun, Rosemary
AU - Kessler, John A.
AU - Espinosa, Horacio D.
N1 - Funding Information:
Research reported in this publication was supported by the National Cancer Institute of the National Institutes of Health under award number U54CA19909 and NIH R21 award number 1R21GM132709-01 .
Publisher Copyright:
© 2023 The Authors
PY - 2023/4
Y1 - 2023/4
N2 - Membrane disruption using Bulk Electroporation (BEP) is a widely used non-viral method for delivering biomolecules 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.
AB - Membrane disruption using Bulk Electroporation (BEP) is a widely used non-viral method for delivering biomolecules 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.
KW - Bulk electroporation
KW - Cell stress response
KW - Intracellular delivery
KW - Localized electroporation
KW - Single cell RNA sequencing
KW - Stem cell
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U2 - 10.1016/j.mtbio.2023.100601
DO - 10.1016/j.mtbio.2023.100601
M3 - Article
C2 - 37063248
AN - SCOPUS:85149850836
SN - 2590-0064
VL - 19
JO - Materials Today Bio
JF - Materials Today Bio
M1 - 100601
ER -