Mukherjee Prithvijit, Peng Chian-Yu, McGuire Tammy, Hwang Jin Wook, Puritz Connor H, Pathak Nibir, Patino Cesar A, Braun Rosemary, Kessler John A, Espinosa Horacio D
Department of Mechanical Engineering, Northwestern University, Evanston, IL, 60208, United States.
Theoretical and Applied Mechanics Program, Northwestern University, Evanston, IL, 60208, United States.
Mater Today Bio. 2023 Mar 4;19:100601. doi: 10.1016/j.mtbio.2023.100601. eCollection 2023 Apr.
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 for therapeutic purposes to 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.
使用体电穿孔（BEP）破坏细胞膜是一种广泛应用的将生物分子递送至细胞的非病毒方法。最近，其微流控对应技术，即局部电穿孔（LEP），已成功应用于从治疗目的到活细胞的时间分析等多种应用。然而，这些过程对基因表达的副作用，可能影响敏感干细胞的生理学，目前尚不清楚。在这里，我们使用单细胞RNA测序（scRNA-seq）来研究BEP和LEP对小鼠神经干细胞（NSC）基因表达的影响。我们的结果表明，与BEP不同，LEP不会导致广泛的细胞死亡或激活可能影响其长期生理学的细胞应激反应途径。此外，我们的证明表明，LEP适用于多天递送方案，因为与BEP相比，它能够更好地保持细胞活力和完整性。
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