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Growth and maturation of small hepatocytes
Author(s) -
MITAKA TOSHIHIRO,
MIZUGUCHI TORU,
SATO FUMIHIKO,
MOCHIZUKI CHIHIRO,
MOCHIZUKI YOHICHI
Publication year - 1998
Publication title -
journal of gastroenterology and hepatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.214
H-Index - 130
eISSN - 1440-1746
pISSN - 0815-9319
DOI - 10.1111/jgh.1998.13.s1.70
Subject(s) - hepatocyte , biology , epidermal growth factor , fetal bovine serum , cell culture , microbiology and biotechnology , cytoplasm , progenitor cell , collagenase , liver cytology , stem cell , andrology , cell , endocrinology , biochemistry , in vitro , medicine , genetics , liver metabolism , enzyme
Proliferation of adult rat hepatocytes is observed in serum‐free Dulbecco's modified Eagle's medium (DMEM) supplemented with 10 mmol/L nicotinamide and 10 ng/mL epidermal growth factor (EGF). The proliferating cells are mainly mononucleate and form small cell colonies surrounded by mature hepatocytes. Although these cells in focal colonies have a less‐differentiated appearance, immunocytochemically and ultrastructurally they possess hepatic characteristics. The size of small hepatocytes is one‐third to half that of mature hepatocytes. Therefore, we call the cells forming a colony, small hepatocytes. The small hepatocytes can be subcultured for several passages. Furthermore, the cells are rich in the supernatant following 50 g centrifugation for 1 min after collagenase liver perfusion. When the cells are cultured in DMEM supplemented with 10% foetal bovine serum, 10 mmol/L nicotinamide, 1 mmol/L ascorbic acid 2‐phosphate, 10 ng/mL EGF and 1% dimethyl sulphoxide, each small hepatocyte can clonally proliferate for more than 3 months. A small hepatocyte divides to form a colony and the number of cells reaches more than 100 within 20 days. With time in culture, cells with a large cytoplasm appear within a colony. They have many mitochondria and large peroxisomes with crystalline nucleoids and are typical, mature hepatocytes. Immunoreactivity to connexin 32 and well‐developed bile canaliculus structures are often observed in the cell‐cell borders. Thus, we suggest that small hepatocytes may be considered to be ‘committed progenitor cells’ that can further differentiate into mature hepatocytes.