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

Prithvijit Mukherjee; Chian Yu Peng; Tammy McGuire; Jin Wook Hwang; Connor H. Puritz; Nibir Pathak; Cesar A. Patino; Rosemary Braun; John A. Kessler; Horacio D. Espinosa

doi:10.1016/j.mtbio.2023.100601

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

Prithvijit Mukherjee, Chian Yu Peng, Tammy McGuire, Jin Wook Hwang, Connor H. Puritz, Nibir Pathak, Cesar A. Patino, Rosemary Braun, John A. Kessler^*, Horacio D. Espinosa^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

7 Scopus citations

Abstract

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.

Original language	English (US)
Article number	100601
Journal	Materials Today Bio
Volume	19
DOIs	https://doi.org/10.1016/j.mtbio.2023.100601
State	Published - Apr 2023

Funding

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 .

Keywords

Bulk electroporation
Cell stress response
Intracellular delivery
Localized electroporation
Single cell RNA sequencing
Stem cell

ASJC Scopus subject areas

Bioengineering
Molecular Biology
Biotechnology
Biomedical Engineering
Cell Biology
Biomaterials

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10.1016/j.mtbio.2023.100601

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title = "Single cell transcriptomics reveals reduced stress response in stem cells manipulated using localized electric fields",

abstract = "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.",

keywords = "Bulk electroporation, Cell stress response, Intracellular delivery, Localized electroporation, Single cell RNA sequencing, Stem cell",

author = "Prithvijit Mukherjee and Peng, {Chian Yu} and Tammy McGuire and Hwang, {Jin Wook} and Puritz, {Connor H.} and Nibir Pathak and Patino, {Cesar A.} and Rosemary Braun and Kessler, {John A.} and Espinosa, {Horacio D.}",

note = "Publisher Copyright: {\textcopyright} 2023 The Authors",

year = "2023",

month = apr,

doi = "10.1016/j.mtbio.2023.100601",

language = "English (US)",

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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.

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

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

Abstract

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Keywords

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