PURPOSE: Bioartificial liver assist devices (BLADs) are 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 thereby limit the use of bioartificial liver therapy. In our lab, fluidized-bed BLADs based on microencapsulated primary porcine hepatocytes have been successfully used to treat liver failure pigs. We detected the risk of PERVs transmission of microencapsulated primary porcine hepatocytes-the key component of fluidized-bed BLADs, to evaluate the biosafety of this device for further clinical applications. METHODS: Microencapsulated primary porcine hepatocytes (cell diameter = 300 μm) were cultured in Dulbecco's modified Eagles medium (DMEM). Microencapsulated cell culture supernatants were collected at 6, 12, 24 and 72 h. HEK-293 were cocultured with these supernatants, and the cocultured cells were harvested every 7 days. RT-PCR was used to detect PERVs transmission. RT-qPCR was used to get the number of virus copies. PK-15 was used as the positive control whereas HepG2 was used as the negative control. RESULTS: PERV was detected in all supernatants, and the viral load of the supernatants increased with time. Moreover, cocultured 293 cells were positive for PERV-specific sequences. CONCLUSION: The kind of fluidized-bed BLADs based on microencapsulated primary porcine hepatocytes have risk of PERVs transmission. Further extensive pre-clinical study focused on biosafety is warranted.
PURPOSE: Bioartificial liver assist devices (BLADs) are expected to bridge liver failurepatients to liver transplantation, but porcine endogenous retroviruses (PERVs) still pose a potential risk in pig-to-human xenotransplantation and thereby limit the use of bioartificial liver therapy. In our lab, fluidized-bed BLADs based on microencapsulated primary porcine hepatocytes have been successfully used to treat liver failurepigs. We detected the risk of PERVs transmission of microencapsulated primary porcine hepatocytes-the key component of fluidized-bed BLADs, to evaluate the biosafety of this device for further clinical applications. METHODS: Microencapsulated primary porcine hepatocytes (cell diameter = 300 μm) were cultured in Dulbecco's modified Eagles medium (DMEM). Microencapsulated cell culture supernatants were collected at 6, 12, 24 and 72 h. HEK-293 were cocultured with these supernatants, and the cocultured cells were harvested every 7 days. RT-PCR was used to detect PERVs transmission. RT-qPCR was used to get the number of virus copies. PK-15 was used as the positive control whereas HepG2 was used as the negative control. RESULTS: PERV was detected in all supernatants, and the viral load of the supernatants increased with time. Moreover, cocultured 293 cells were positive for PERV-specific sequences. CONCLUSION: The kind of fluidized-bed BLADs based on microencapsulated primary porcine hepatocytes have risk of PERVs transmission. Further extensive pre-clinical study focused on biosafety is warranted.
Authors: A J Ellis; R D Hughes; J A Wendon; J Dunne; P G Langley; J H Kelly; G T Gislason; N L Sussman; R Williams Journal: Hepatology Date: 1996-12 Impact factor: 17.425
Authors: F D Watanabe; C J Mullon; W R Hewitt; N Arkadopoulos; E Kahaku; S Eguchi; T Khalili; W Arnaout; C R Shackleton; J Rozga; B Solomon; A A Demetriou Journal: Ann Surg Date: 1997-05 Impact factor: 12.969
Authors: G V Mazariegos; D J Kramer; R C Lopez; A O Shakil; A J Rosenbloom; M DeVera; M Giraldo; T A Grogan; Y Zhu; M L Fulmer; B P Amiot; J F Patzer Journal: ASAIO J Date: 2001 Sep-Oct Impact factor: 2.872