Isolation And Identification Of Adipose Stem Cell Exosomes And The Study Of Anti-hepatocellular Carcinoma Efficacy Of Its Delivery Of Curcumin In Vitro

Cancer has become an important cause of death for residents in China,and currently chemotherapy is still one of the most common means.Most small molecule anticancer drugs compounds have high lipophilicity and low water solubility,resulting in low bioavailability,which is often regarded as a key obstacle to the development and clinical application of these anti-cancer drugs.Current studies have found that mesenchymal stem cell exosomes have similar therapeutic effects as stem cells,and have the advantages of easy freezing,transportation,quantification,and transplantation.Because of their many biologically active molecules that can be passed between cells,they are considered to have good potential as a drug delivery vehicle.However,the study found that differences in the means of exosomes isolation may significantly affect the function of exosomes,and it is unclear whether the biological effects of adipose-derived mesenchymal stem cell-derived exosomes on liver cancer,nor whether there is a synergistic effect between exosomes and small molecule anticancer drugs.Therefore,the study found that:(1)ADSCs exosomes separated by improved sequential filtration method,the particle size is mainly distributed in 30 ~ 150 nm,has a membrane structure,positively expresses TSG101,CD63?CD9,GADPH,and negatively expresses Calnexin.After the procedure of isolation in vitro,ADSCs exosomes still has the ability to be taken up by cells and has biological activity.The exosomes of ADSCs obtained by this method inhibits the cell activity and migration ability of Hep G2,and enhances the cell activity of THLE-2.(2)At 300?g / m L exosome concentration and 50?g / m L astaxanthin concentration,the drug loading efficiency: electroporation method> freeze-thaw cycle method> co-incubation method> saponin permeabilization method.The application of electroporation to the loading of exosomes has advantages.(3)ADSCs exosomes isolated by the improved ultrafiltration method could inhibit the activity of Hep G2(P <0.05)and its migration ability(P <0.05)and enhance the activity of THLE-2 within 24 h.(P <0.05)and had no significant effect on its migration ability.The simple mixing of ADSCs exosomes and curcumin inhibited the migration ability of TLHE-2(P<0.01).The curcumin-encapsulated exosomes prepared by electroporation can inhibit the activity of Hep G2(P<0.01)and its migration ability(P <0.01).The drug loading process by electroporation enhances the inhibition of Hep G2 activity and its migration ability,and weakened the promotion of THLE-2 migration ability,compared to the mixture of exosomes and curcumin.The results suggest that the ADSCs exosomes isolated by this method have the prospect of application in the treatment of liver cancer,and can be used for drug delivery to enhance its anti-liver cancer effect.