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Fig. 1. Development and maturation of small-hepatocyte colonies in culture. (A) A small-hepatocyte colony after 1 week of culture. (B) A small-hepatocyte colony after 7 weeks of culture. Notice the hexagonal shape of most cells, indicating differentiation of small hepatocytes into mature hepatocytes. (C) Phase-contrast microscopy picture showing a clearly demarcated colony of cells appearing as mature hepatocytes. Cells were cultured for 9 weeks and then fixed with 4% paraformaldehyde in the presence of Triton X-100 and subsequently processed for immunofluorescence localization of Mrp2. (D) Immunofluorescence localization by laser-scanning microscopy indicated expression of Mrp2 in structures resembling bile canaliculi. Scale bars, 40 µm.





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