Gene Delivery via Octadecylamine-Based Nanoparticles for iPSC Generation from CCD1072-SK Fibroblast Cells

Area of Science:

• Biotechnology
• Stem Cell Biology
• Nanomedicine

Background:

• Induced pluripotent stem cells (iPSCs) hold promise for regenerative medicine.
• Traditional viral vectors for iPSC reprogramming pose safety concerns.
• Development of non-viral delivery systems is crucial for efficient and safe iPSC generation.

Purpose of the Study:

• To develop and characterize novel octadecylamine-based solid lipid nanoparticles (OCTNPs) as a non-viral vector for iPSC reprogramming.
• To evaluate the efficiency of OCTNPs in delivering reprogramming factors to human fibroblast cells.
• To confirm the pluripotency of generated iPSCs and their differentiation potential.

Main Methods:

• Synthesis and characterization of OCTNPs (size, zeta potential).
• Transfection of human CCD1072-SK fibroblast cells with reprogramming factors using OCTNPs.
• Characterization of iPSCs using immunofluorescence, flow cytometry, RT-qPCR, and alkaline phosphatase assay.
• Differentiation of iPSCs into cardiomyocyte-like cells using 5-azacytidine.

Main Results:

• OCTNPs were successfully synthesized with an average size of 178.9 nm and a positive zeta potential of 22.8 mV.
• OCTNPs demonstrated high transfection efficiency (82.0%) for delivering reprogramming factors.
• Generated iPSCs expressed key pluripotency markers (OCT4, SOX2, KLF4) and showed alkaline phosphatase activity.
• iPSCs were successfully differentiated into cardiomyocyte-like cells.

Conclusions:

• OCTNPs represent a novel and effective non-viral vector for iPSC reprogramming.
• This approach offers a safer alternative to viral vectors, minimizing risks in stem cell applications.
• The findings support the potential of OCTNPs in advancing biotechnological applications, including tissue engineering and personalized medicine.