Stable Hepatocyte Engraftment after Hepatocyte Transplantation Using Biodegradable Injectable Polymer

Choi, Dong Ho; Roh, Eun; Kim, Han Joon; Lee, Kyeong Geun; Paik, Seung Sam; Lee, Sang Hun; Lee, Yong Sung; Paik, Doo Jin; Kim, Byung Soo; Lee, Kwang Soo

Abstract

PURPOSE: Whole liver transplantation, an effective therapy for many inherited and acquired hepatic disorders, has limitations including donor shortage and fatal surgical complications. Hepatocyte transplantation, which is simpler and less expensive than whole liver transplantation, allows the use of living related donors, permits the use of a single donor organ for multiple recipients, and makes possible the cryopreservation of hepatocytes for future use. However, choosing a proper scaffold for hepatocytes hampers wide use of hepatocyte transplantation. We performed hepatocyte transplantation using biodegradable injectable polymers, fabricated form poly (lactide-co-glycolide) (PLGA) microspheres, as scaffolds to evaluate their effectiveness.
METHODS
Female, five week old FVB mice, were prepared for donors, and four male, five week old nude mice, were used for recipients. Liver cells were isolated from FVB donors. The cell viability exceeded 95% as assessed by trypan blue exclusion method. For three nude mice, 2X10(6) cells resuspended in 200micro liter medium were mixed with 200micro liter PLGA microspheres, and were injected into the peritoneal cavity of each mouse. One nude mouse was transplanted with 2X10(6) cells resuspended in 200micro liter medium only, and it served as a negative control. Specimens were retrieved at one week, and histological and immunohistochemical analyses were performed.
RESULTS
In the negative control, all transplanted hepatocytes disappeared at one week. In mice transplanted both microspheres and hepatocytes, conglomerates, which contained hepatocytes, were observed in the peritoneal cavity, The hepatocytes were identified by H and E staining and immunohistochemistry using anti- hepatocyte antibody.
CONCLUSION
In this preliminary study, stable hepatocyte engraftment was achieved in hepatocyte transplantation with PLGA microspheres, but not in hepatocyte transplantation only. More studies on comparison between sponge-type scaffolds and injectable scaffolds would be necessary. Improvement on both initial vascularization and proliferation of transplanted hepatocytes is a target of our future work.