Generation of murine induced pluripotent stem cells by using high-density distributed electrodes network

Area of Science:

• Biotechnology
• Cell Biology
• Gene Delivery

Background:

• Traditional gene delivery methods often rely on viral vectors, which can pose safety concerns.
• Non-viral methods are sought after for safer and more efficient gene delivery, particularly for generating induced pluripotent stem cells (iPSCs).

Purpose of the Study:

• To develop and validate a novel high-density electrodes network (HDEN) device for robust gene transfection in murine primary cells.
• To optimize the generation of iPSCs using non-viral gene delivery with the HDEN device.

Main Methods:

• Utilized a high-density electrodes network (HDEN) for gene transfection in a murine primary cell model.
• Transfected cells with plasmid DNA encoding fluorescent proteins and iPSC factors.
• Assessed cell viability, transgene expression levels, and transfection efficiency.

Main Results:

• Achieved high cell viability (>95%) post-transfection.
• Demonstrated successful and steady transgene expression, including fluorescent proteins and iPSC factors.
• Reported transfection rates of 42.4% (single) and 24.5% (triple) with low voltage (<180 V) and low cell numbers (<1.5 × 10^4).
• Observed over 10-fold higher iPSC factor gene expression compared to previous studies.

Conclusions:

• The HDEN device offers a promising, efficient, and safe non-viral method for gene transfection.
• HDEN facilitates optimized reprogramming efficiency for generating murine iPSCs.
• This technology presents a feasible alternative to viral transfection methods for iPSC generation.