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

Prithvijit Mukherjee1,2, Chian-Yu Peng3, Tammy McGuire3

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Localized Electroporation (LEP) offers a gentler alternative to Bulk Electroporation (BEP) for cell manipulation. LEP preserves neural stem cell viability and gene expression, unlike BEP, making it suitable for sensitive cell applications.

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Area of Science:

  • Biotechnology
  • Cell Biology
  • Genomics

Background:

  • Electroporation is a key technique for delivering molecules into cells.
  • Localized Electroporation (LEP) is a newer microfluidic method with potential applications in cell reprogramming and sampling.
  • The impact of electroporation methods on sensitive stem cell gene expression is not fully understood.

Purpose of the Study:

  • To investigate the effects of Bulk Electroporation (BEP) and Localized Electroporation (LEP) on murine neural stem cell (NSC) gene expression.
  • To compare the cellular impact, including viability and stress responses, of BEP and LEP.

Main Methods:

  • Single cell RNA sequencing (scRNA-seq) was employed to analyze gene expression profiles.
  • Murine neural stem cells (NSCs) were subjected to both BEP and LEP.

Main Results:

  • LEP did not induce significant cell death or activate stress response pathways in NSCs, unlike BEP.
  • LEP demonstrated superior preservation of cell viability and integrity compared to BEP.
  • LEP is suitable for multi-day cell delivery protocols.

Conclusions:

  • LEP is a less disruptive method than BEP for genetic manipulation of neural stem cells.
  • LEP preserves NSC physiology and viability, making it advantageous for therapeutic and research applications.
  • scRNA-seq analysis provides critical insights into the molecular effects of electroporation techniques on stem cells.