Exosomes facilitate mRNA and siRNA delivery using cationic liposomes 2X3-DOPE to rat heart mesenchymal cells <i>in vitro</i>

作者：Olesya Dovbysh, Vera V. Vysochinskaya, Nina V. Gavrilova, Pavel M. Docshin, Ekaterina Nikitina, Aleksandr S. Klochev, Е. А. Елпаева, Olga A. Dobrovolskaya, Elena V. Shmendel, М. А. Маслов, Yana Zabrodskaya · 发表于：Medical academic journal · 年份：2025 · DOI：10.17816/maj641910 · 研究领域：Extracellular vesicles in disease、RNA Interference and Gene Delivery、MicroRNA in disease regulation

BACKGROUND: The delivery of nucleic acids to mesenchymal stem cells, which are used as model objects in in vitro experiments or as therapeutic agents in regenerative medicine and oncology, is an actively developing area of research. Existing non-viral delivery systems either have low effectiveness or highly toxic to mesenchymal stem cells. Therefore, the development of new carriers has become an urgent priority. AIM: To demonstrate the feasibility of delivering model messenger RNA and small interfering RNA to rat heart mesenchymal stem cells (MSCs) in vitro using original cationic liposomes 2X3-DOPE (1:3 molar ratio) and to evaluate the influence of exosomes incorporated into hybrid nanoparticles with 2X3-DOPE on the efficiency of RNA delivery. METHODS: Exosomes were isolated using a standard ultracentrifugation technique followed by characterization of the obtained vesicles through Western blotting, transmission electron microscopy, atomic force microscopy, and hydrodynamic diameter measurement using dynamic light scattering. Small interfering RNA was chemically synthesized; whereas messenger RNA was obtained by in vitro transcription. Complexes of liposomes or hybrid nanoparticles with RNA were prepared by mixing; the properties of the resulting particles were assessed using dynamic light scattering and atomic force microscopy. To evaluate the efficiency of RNA delivery to rat heart mesenchymal stem cells derived from both healthy and ischemic myocardium, we used fluorescen...