Preparation, characterisation and cell transfection of cationic liposomes in gene therapy

Cationic lipid-mediated gene transfer is one of the most commonly used non-viralvectors. It has been shown to be a safe and effective carrier. However, its use ingene delivery was hampered by its low transfection efficiency and stability.DOTAP, DOPE, cholesterol (CHO) and carboxymethyl-β-cyclodextrin (CD) wereused to prepare cationic liposomes. Cationic liposomes were prepared using both,thin film hydration and a microfluidic method. Formulation stability wasevaluated using liposome size, zeta potential and polydispersity index (PDI).Promega QuantiFluor® ONE dsDNA System was used to investigate theencapsulation efficiency. COS7 and SH-SY5Y cell lines were used to determinetransfection efficiency. Results show that carboxymethyl-β-cyclodextrin increasedencapsulation efficiency by 15.5% and 8% using NanoAssemblr® and rotaryevaporator, respectively compared to liposomes without CD. The addition ofcarboxymethyl-β-cyclodextrin to cationic liposomes resulted in an increase intransfection efficiency in both cell lines.B