A new source of hepatocytes for transplantation.
BACKGROUND
The most effective treatment for acute or chronic liver failure is orthotopic liver transplantation. Worldwide there is a shortage of organs for transplantation. This shortage has called for research into new treatments for management of patients with liver failure. One such treatment is hepatocyte transplantation. During liver resections considerable amounts of normal liver are unavoidably resected. We aim to harvest these hepatocytes and to filter the tumor cells from them to provide a source for transplantation.


METHODS
After liver resection, the largest vessel at the resected liver edge was identified and cannulated. Seglen's two-stage technique of perfusing the liver with EDTA and collagenase was performed to harvest the hepatocytes. Ep-CAM Ags are consistently present on the surface of epithelial cells and in particular in colorectal cancer cells. Therefore, MOC31 antibodies (selective Abs for Ep-CAM) attached to magnetic beads were used to target the tumor cells. These tumor cells are selectively removed using a magnet. CEA staining was then used to ensure the hepatocyte collection was tumor cell free. Five million hepatocytes were rosetted with one million HT29 CRC cells to assess the immunomagnetic filtration technique.


RESULTS
The hepatocyte harvesting resulted in 864,000 viable hepatocytes to be harvested per gram of liver. Histochemical staining using CEA demonstrated 75% of the HT29 cells in the hepatocyte collection were removed after one use of magnetic beads.


CONCLUSIONS
We have demonstrated the successful initial stages of harvesting tumor-free hepatocytes from liver resected for malignancy.
Haghighi
K S
KS
UNSW Department of Surgery, St George Hospital, Sydney, Australia.
Woon
W W L
WW
Akhter
J
J
Marr
P J
PJ
Bolton
E
E
Riordan
S
S
Morris
D L
DL
D000368:Aged; D002272:Carcinoembryonic Antigen / Q000032:analysis; D015179:Colorectal Neoplasms / Q000473:pathology; D005260:Female; D006498:Hepatectomy / Q000379:methods; D022781:Hepatocytes / Q000473:pathology / Q000637:transplantation*; D006801:Humans; D018189:Immunomagnetic Separation; D008113:Liver Neoplasms / Q000473:pathology / Q000556:secondary* / Q000601:surgery*; D008297:Male; D008875:Middle Aged; D020858:Tissue and Organ Harvesting / Q000379:methods
D002272:Carcinoembryonic Antigen