期刊
EUROPEAN JOURNAL OF MEDICAL RESEARCH
卷 17, 期 -, 页码 -出版社
BMC
DOI: 10.1186/2047-783X-17-13
关键词
Microbiological safety; Porcine endogenous retrovirus; Bio-artificial liver; Porcine hepatocyte; Mesenchymal stem cells
资金
- Natural Science Foundation of Jiangsu Province [BK2006008]
- foundation of Medical Center of Jiangsu Province [ZX200605]
Background: Our institute has developed a novel bio-artificial liver (BAL) support system, based on a multi-layer radial-flow bioreactor carrying porcine hepatocytes and mesenchymal stem cells. It has been shown that porcine hepatocytes are capable of carrying infectious porcine endogenous retroviruses (PERVs) into human cells, thus the microbiological safety of any such system must be confirmed before clinical trials can be performed. In this study, we focused on assessing the status of PERV infection in beagles treated with the novel BAL. Methods: Five normal beagles were treated with the novel BAL for 6 hours. The study was conducted for 6 months, during which plasma was collected from the BAL and whole blood from the beagles at regular intervals. DNA and RNA in both the collected peripheral blood mononuclear cells (PBMCs) and plasma samples were extracted for conventional PCR and reverse transcriptase (RT)-PCR with PERV-specific primers and the porcine-specific primer Sus scrofa cytochrome B. Meanwhile, the RT activity and the in vitro infectivity of the plasma were measured. Results: Positive PERV RNA and RT activity were detected only in the plasma samples taken from the third circuit of the BAL system. All other samples including PBMCs and other plasma samples were negative for PERV RNA, PERV DNA, and RT activity. In the in vitro infection experiment, no infection was found in HEK293 cells treated with plasma. Conclusions: No infective PERV was detected in the experimental animals, thus the novel BAL had a reliable microbiological safety profile.
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