BACKGROUND/AIMS: This study was designed to clarify the fine structures of the hepatocytes and mesencymal tissues in chronic hepatitis according to severity. METHOD: For the purpose of elucidating the ultrastructural characteristics of mesenchymal tissues, liver biopsy specimens were studied by light and electron microscopy in 20 patients with chronic hepatitis. RESULTS: 1) Hepatocytes in mesenchymal tissues were thought to be in the stage of regenerated or degenerated process. 2) Regenerating nodules were surrounded by a basement membrane-like materials in the space of Disse. 3) In the widened Disse space the deposition of collagen fiber bundles and increased numbers of hepatic stellate cells in necrotic area were observed. 4) In necrotic areas, hepatic mesenchymal cell response including an increase of collagen fibers and fibroblast, angiogenesis, and a proliferation of bile ductules were also observed. CONCLUSIONS: These observations suggest that the fibrosis in severe chronic hepatitis was accompanied by the mesenchymal response including the proliferation of hepatic stellate cells, fibroblasts, capillarization of Disse space, and mesenchymal proliferation. Finally, this fibrosis observed electron microscopically may be a cause of functional hepatic failure.
Adult ; English Abstract ; Female ; Hepatitis, Chronic/*pathology/virology ; Hepatitis, Viral, Human/*pathology ; Hepatocytes/ultrastructure ; Human ; Liver/*ultrasonography ; Male ; Mesoderm/ultrastructure ; Microscopy, Electron ; Middle Aged

Adult human bone marrow-derived mesenchymal stem cells (MSCs) were cultured on microcarriers in spinner flasks and were compared with those in conventional culture in 12-well plates. For the production of adherently growing MSCs, macroporous CultiSpher G gelatin microcarriers were used in concentration of 1 g/L. The cells were seeded in a density of 5 x 10(4) cells/ml in both spinner culture and conventional stationary culture. The result showed that after 7 days of cultivation a maximum viable cell concentration of 5.15 x 10(5) cells/ml was obtained in spinner culture. Whereas the cell density increased to a maximum of 1.675 x 10(5) cells/ml on day 5 in conventional stationary culture. Lactate was produced up to 12.06 mmol/L in spinner culture and up to 13.10 mmol/L in stationary culture, and glucose was consumed up to 7.38 mmol/L and 5.37 mmol/L respectively. The average lactate yield on glucose consumption in spinner culture was only 1.63, lower than that in stationary culture 2.44. This indicated that the energy metabolism in spinner culture was significantly more efficient than that in conventional culture. After spinner culture for 12 days, the MSCs maintain the characteristics of stem cells. It is concluded that the microcarrier culture system is a suitable way to expand the seeding cells for tissue engineering.
Adult ; Antigens, CD ; analysis ; Bone Marrow Cells ; cytology ; metabolism ; ultrastructure ; Cell Count ; Cell Culture Techniques ; instrumentation ; methods ; Cell Division ; Cell Survival ; Flow Cytometry ; Glucose ; metabolism ; HLA-DR Antigens ; analysis ; Humans ; Lactates ; metabolism ; Mesoderm ; cytology ; metabolism ; ultrastructure ; Microscopy, Electron ; Microscopy, Electron, Scanning ; Stem Cells ; cytology ; metabolism ; ultrastructure ; Time Factors