Preparation of stearamide-based nanoparticle systems and using in vitro gene transfection of fibroblast cells

Induced pluripotent stem cells (IPSC) are preferred as an alternative source in regenerative medicine, disease modeling and drug screening thanks to their unique properties compared to other stem cells. As seen from the previous studies in the literature, most of the vector systems used for transfection of somatic cells to obtain IPSC are viral-based. The application of IPSC derived via viral vector systems, are limited due to immune response and tumor formation. The aim of this study is to develop a non viral vector system for transfection of Yamanaka factors and to obtain IPSC. In this study, SLNP synthesis conditons in order to obtain high transfection efficacy were studied. In this study, cationic stearamide cetylpyridium chloride lipid nanoparticles (SCLN) which are in solid lipid nanoparticle class are prepared via solvent diffusion method. The obtained NP system is going to be used for the delivery of plasmid encoding Oct3/4, Sox2, Klf4, v-Myc and GFP to fibroblast cell line. Optimization studies on zeta potential and particle size have been performed in order to achieve low cytotoxicity and to eliminate macrophage clearance. Cytotoxicity studies for the plasmid/stearamide nanoparticle conjugate have been performed via L929 cell line. Later, transfection of fibroblast cells has been carried out by using plasmid stearamide nanoparticle complex and the transfection efficiency was observed by following GFP expression with the inverted fluorescent microscope for 4 days. The expression of the Oct3 / 4, Sox2, Klf4 and v-Myc genes which are IPS cell markers were determined by RT-qPCR. The stem cells are promisind cells and seem as a solution for serious problems encountered in medical fields and bioengineering such as wound healing in diabetes, cardiological diseases and nerve-tissue engineering. Keywords: reprogrammed cells, stem cell, cationic nanoparticle, stearamide