Fluidized-bed Bioartificial Liver Assist Devices (BLADs) Based On Microencapsulated Primary Porcine Hepatocytes Have Risk Of PERVs Transmission

Background:bioartificial liver assist devices(BLADs) is expected to bridge liver failure patients to liver transplantation, but porcine endogenous retroviruses (PERVs) still pose a potential risk in pig-to-human xenotransplantation and bioartificial liver therapy. In our lab, fluidized-bed bioartificial liver assist devices (BLADs) based on Microencapsulated primary porcine hepatocytes have been successfully used to treated liver failure pigs, we detected the risk of PERVs transmission of microencapsulated primary porcine hepatocytes, which is the key component of fluidized-bed bioartificial liver assist devices, to evaluate the biosafety of this device for further use.Methods:Microencapsulated primary porcine hepatocytes (cell diameter=300um) were cultured in DMEM. We collected 6h,12h,24h,72h microencapsulated cells'culture supernatants. HEK-293 were cocultured with these supernatants, and the cocultured cells were collected every seven days. RT-PCR were used to investigate the PERVs transmission, RT-qPCR to measure the quantities of virus load. PK-15 was taken as positive control, and HepG2 as negative control.Results:PERV was detected in all supernatants, and the viral load of supernatants increased with time. Moreover, infected 293 cells were positive for PERV-specific sequences.Conclusion:The kind of fluidized-bed bioartificial liver assist devices (BLADs) based on Microencapsulated primary porcine hepatocytes seems have risk of PERVs transmission; extensive pre-clinical study focused on biosafety should be made.