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    Enhancing transfection efficiency of primary cell lines using different terminated PBAE structures without endcapping reaction
    (Springer, 2025) Demir, Irmak; Celik, Sibel Kucukertugrul; Bal, Kevser; Kaplan, Ozlem; Senturk, Sema; Demir, Kamber; Gok, Mehmet Koray
    Gene therapy holds promise for a wide range of diseases, including Alzheimer's, diabetes, and cancer, and requires the efficient transfer of nucleic acids into cells. However, transfection in primary cells is still problematic and requires the development of new transfection agents. Poly (beta-amino ester) (PBAE) has attracted great attention in transfection research due to their low toxicity, high gene loading capacity, endosomal escape ability, and biodegradability properties. In this study, two new PBAEs with different molecular weights are synthesized that could provide high viability and transfection efficiency in primary cells. They are characterized using FTIR and 1H NMR analysis. GPC-SEC system is also used to calculate the average molecular weight (Mw) and polydispersity index. PBAE nanoparticle preparation is carried out using the nanoprecipitation technique. The gene loading capacity, protective ability against nuclease degradation, and proton buffering capacity of nanoparticles are determined. Additionally, the morphology of PBAEA:pEGFN1 complexes was investigated by STEM analysis. Finally, their cytotoxicity and transfection efficiency in primary ovine fibroblast (POF) cells are also investigated. The results reveal that the new PBAE with higher Mw achieves quite high transfection efficiency of about 87% and did not show any cytotoxic effects on these cells. These findings suggest that PBAE is a promising option to achieve high transfection efficiency in primary cells.