Ken Yaegaki
Nippon Dental University, Japan
Title: Hepatocytes produced from human tooth pulp into swine with cirrhosis: Two transplantation with time interval
Biography
Biography: Ken Yaegaki
Abstract
We treated the liver cirrhosis by two step transplantations of hepatocyte produced from human exfoliated deciduous tooth (SHED) into the swine with cirrhosis. SHED at 3 passages was separated by magnetic sorting with CD117 antibody. For hepatic differentiation, CD117+ SHED were grown in DMEM supplemented with insulin-transferrin-selenium-x (ITS-x), embryo-tropic-factors(ETF) and hepatocyte-growth-factor (HGF) for 5days: IMDM supplemented with ITS-x, ETF, HGF, dexamethasone and oncostatin for another 11 days. F344-Nude rats were employed for this study. Carbon tetrachloride (CCl4) was administrated by intraperitoneal injection for 15 weeks to induce cirrhosis. Hepatocyte-like-cells (2 x 106 cells/ animal) suspended in Hank`s Balanced Salt Solution were transplanted into the spleen. The vehicle was injected to the positive control group. Non-cirrhosis-models were used as negative control group. Animals were sacrificed 4 weeks after the transplantation. Then 5 weeks later the sound transplantation was carried out, then the swine were euthanased. Immunocytochemistry observation of the hepatically differentiated cells strongly demonstrated positive staining for albumin, IGF-1, α-feto-protein, HNF4α and CPS-1. The histopathological analysis, HE and Masson’s trichrome staining, indicated a significant decrease of fibrous tissue in the transplantation group with comparing to the positive control group. Healthy liver tissues were recovered by the transplantation. Moreover, serological test results revealed significance differences between the groups. Serum ALT levels of the test group dramatically decreased to one third compared to the positive control group. Activities of albumin, bilirubin, BUN, HA levels were also recovered. By only once transplantation albumin value was improved. but two steps showed much better improvement. The two steps transplantations of hepatocyte-like cells from human tooth transplanted into the liver with severe failure demonstrated their capacity to preform positively because of drastic decreasing fibrous tissues. Together, these findings suggested that two step transplantation is a future potential protocol for treating chronic liver injuries such as cirrhosis.