Background: It is critical issue for clinical that liver failure after large hepatectomy with cholangiocellular carcinoma. This time, we paid attention to this recected liver. This recected liver have a large quantity of normal hepatocytes with carcinoma cell. If completely divited two group using magnet sorting by tumor Ab, these cell is able to transplant. It is phenomenal finding if it can be possible to auto-transplanted into the patient only normal hepatocyte.
Methods: Hepatocyte isolation from wild type rat(Lewis Rat) were performed by manual enzymatic perfusion with collagenase. Those hepatocyte were mixed with HepG2, which is hepatocellular carcinoma cell line, equally. To remove HepG2 cells, they were sorted by magnetic method with Anti-Telomerase reverse transcriptase antibody which is specific antibody for HepG2 using DynaMag™-15 Magnet.
For evaluation that separation was completed, we performed flow cytometory by Anti-Telomerase reverse transcriptase antibody and cell transplantation into Nagase rat which is non-albumic rat, and measured serum albumin level(Day3,7,10,14) using ELISA-based turbidimetric immunoassay and histological findings with albumin staining. we also transpanted those cells into nude mice for tumorigenesis
Result: Albumin levels significantly increased in the magnetic-sorting group than in control group. Simultaneously, the spleen in which these hepatocytes were transplanted could be used to observe hepatocytes; the cells were transplanted 14 days later, and the magnetic-sorting group had significantly higher hepatocyte levels than the control group. And also we could not see any tumorigenesis in nude mice during three month.
Conclusion: We developed an effective technique of hepatocyte isolation for transplantation. As a result, it was believed that this method could not only improve cell transplant effect but also store cells safely. These results demonstrate the usefulness of hepatocytes for cell transplantation.