Experimental research for treatment of liver failure with hepatic tissue stem cell (SP cell)

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1. Hepatic differentiation induction method in vitroIn an effort to reconstruct the hepatocyte function and cellular polarity normally found in the liver, adult rat hepatocytes were sandwiched between two layers of hydrated collagen matrix. Functionally, sandwiched hepatocytes maintained the secretion of albumin, the expression of liver specific proteins and the distribution of actin filaments, whereas the cells cultured on single layer of collagen gel decreased the albumin secretion, the liver specific proteins and showed abnormal formation of actin stress fibers and cell spreading. Overlaying a second layer of collagen gels on the hepatocytes that had been cultured on a single gel reversed the cell spreading, reduced actin stress fibers and recovered the liver specific protein expressions.Bone marrow cells could differentiate into hepatocytes when they co-cultured with primary cultured hepatocytes in collagen gel sandwich indicating that hepatocytes cultured in collagen gel sandwich was functionally good enough to induce differentiation of bone marrow cell into hepatocytes.2. SP cellsWe could successfully induce SP cells derived from liver and bone marrow transdifferentiation into hepatocytes in vitro and in vivo. Splenic SP cells have a remarkable capacity for hepatic differentiation, as they were co-cultured with primary cultured hepatocytes in collagen gel. It is our great surprise that the prevalence of SP cells in spleen is 5 to 10 times higher than in bone marrow. Our results indicated that splenic-SP cells might have the potential to transdifferentiate into hepatocytes. Since the prevalence of SP cells in spleen is much higher than in bone marrow, total amount of SP cells could be much larger. Thus the spleen could have the highest prevalence of SP cells and spleen could provide an alternative source of stem cell for stem cell transplantation into liver.