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Primary cultures of rat hepatocytes and hepatic stellate cells were produced from 7-d-old neonatal livers by incubation of the liver in 2.5% type IV collagenase for 8 h at 37 degrees C. Hepatocytes and stellate cells were purified from the digested cell suspension by centrifugation in Percoll density gradients, stained with cytochemical and immunocytochemical stains, and characterized by light and electron microscopy. Hepatocytes were identified by their major morphologic features and their uniform staining with an antibody to acetylated cytokeratin (F8). Hepatic stellate cells were identified by their flattened morphology and vacuolated cytoplasm. Both cell types could be maintained in culture for greater than 15 d. At the electron microscopic level, the hepatocytes displayed abundant rough endoplasmic reticulum, well-developed mitochondria with tubular cristae, and large polyribosomes. Hepatic stellate cells contained abundant mitochondria and rough endoplasmic reticulum, and their cytoplasmic matrix showed many lipid droplets and the Golgi complex. The presence of both hepatocytes and stellate cells in the same culture demonstrated the following: the hepatocytes appeared to produce large amounts of collagen, which was broken down by the stellate cells, some of which became lipid-laden. A model of these interactions is shown.Girls masturbating in their chair and on the floor. Dick smacks and hands
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