MAGNETOFECTION OF PLASMID DNA WITH MESSENGER GENE INTO MELANOMA CELLS B16F1 CULTIVATED IN VITRO WITH SUPERPARAMEGNETIC NANOPARTICLES IN THE PRESENCE OF OUTER MAGNETIC FIELD

摘要

The invention refers to a magnetofection of plasmid DNA (pDNA) with the messenger genome into B16F1 melanoma cells grown in vitro, with superparamagnetic nanoparticles in the presence of an external magneticfield, which includes the process of precipitation of aqueous solutions of sulfates, chlorides and nitrates of the metal ions Fe2+, Fe3+, Mn2+, Zn2+ with water solutions of alkaline precipitating reagents NaOH and NH4OH at pH values in the range between 7 and 13, in the temperature range between 20 degrees C and 100 degrees C, and the dwell time at the maximum temperature from 1 to 120 minutes, the process of surface treatment of synthesized oxide nanoparticles with an aqueous solution of acid stabilization reagent HNO3, anionic hydrophilic polymers (sodium salt of polyacrylic acid, polyactic acid polyglycolic acid, polyacrylic maleinic acid), cationic hydrophilic polymers (polyethyleneimines (PEI), poly lysine, chitin, aminated dextran), at pH values in range between 2 and 11, in the temperature range between 20 degrees C and 50 degrees C, and the dwell time at a maximum temperature of 10 seconds to 240 minutes, the process of binding of an aqueous solution of PEI and pDNA to the pre-sterilized superparamagnetic nanoparticles by filtration through a filter with pore size 0.22 micrometer, the process of transfection of superparamagnetic nanoparticles complexes, PEI, pDNA into the B16F1 melanoma cells grown in vitro in AMEM culture medium with fetal calf serum, L-glutamine, gentamycin and penicillin on transparent microtiter plates using external magnetic field with magnetic flux density 245 mT and gradient magnetic field 40 T / m, and adapted to delivery conditions of genetic material and active substances into the cells grown in vitro for various biomedical and biotechnological applications.