RESUMEN
Liver cell isolation and transplantation have been successfully performed in animal models and in humans. However, lack of initial engraftment due to cell death is a major roadblock to achieving clinical significance. Apoptosis was recently identified as an important cause of freshly isolated and banked hepatocyte cell death [Cell Transplant. 10 (2001) 59]. Pathways involving detachment-induced apoptosis (anoikis) are well characterized in other cell types. Loss of cell anchorage occurs during the hepatocyte isolation procedure prior to cell transplantation, but little is known about the role of this pathway in the survival of isolated hepatocytes. We report early occurrence of anoikis in primary mouse hepatocytes cultured under detached conditions on glass plates as compared to under attached conditions on plastic plates. Apoptosis in detached cells was determined using complementary techniques (DNA laddering, cell death ELISA assay, TUNEL assay and morphological analysis) and was detected as early as 15 min after culture under detached conditions. Further analysis of the mechanisms inducing apoptosis during liver cell isolation and transplantation and of ways to prevent them could lead to improved clinical protocols of liver cell therapies.
