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Objective: To investigate the nature of the restoration of cell polarity and bile excretion function in Sandwich-cultured hepatocytes. Methods: Freshly isolated hepatocytes from male Sprague-Dawley rats were cultured in a double layer collagen gel Sandwich configuration. Morphological changes were observed under a inverted microscope. The domain specific membrane associated protein DPPⅣwas tested by immunofluorescence. and the bile excretion function was determined by using fluorescein diacetate. Hepatocytes cultured on a single layer of collagen gel were taken as control. Results: Adult rat hepatocytes cultured in a double layer collagen gel sandwich configuration regained its morphological and functional polarity and maintained polygonal morphology for at least 4 weeks. Immunofluorescence studies using antibodies against DPPⅣshowed polarity restoration as early as 48 h. After cultured in the double layer collagen gel Sandwich configuration for 96 h the hepatocytes began to excrete bile; while hepatocytes cultured on a single layer collagen gel had no bile excretion. Conclusion: Hepatocytes cultured in a double layer collagen gel Sandwich configuration are able to regain their morphological and functional polarity given certain conditions. Hepaotcyte culture is a useful tool for the study of polarity restoration.
Objective: To investigate the nature of the restoration of cell polarity and bile excretion function in Sandwich-cultured hepatocytes. Methods: Freshly isolated hepatocytes from male Sprague-Dawley rats were cultured in a double layer collagen gel Sandwich configuration. Morphological changes were observed under a inverted microscope. The domain specific membrane associated protein DPPⅣwas tested by immunofluorescence. and the bile excretion function was determined by using fluorescein diacetate. Hepatocytes cultured on a single layer of collagen gel were taken as control. Immunofluorescence studies using antibodies against DPP IVshowed polarity restoration as early as 48 h. After cultured in the double layer collagen gel Sandwich configuration for 96 h the hepatocytes began to excret while hepatocytes cultured on a single layer collagen gel had no bile excretion. Conclusion: Hepatocytes cultured in a double layer collagen gel Sandwich configuration are able to regain their morphological and functional polarity given certain conditions. Hepaotcyte culture is a useful tool for the study of polarity restoration.