A simple method for the isolation of human Ito cells (Fat-storing cells)

• Published in: Kanzo Vol. 32; no. 2; pp. 169 - 174
• Main Authors: NAKANO, Hiroshi, NAKABAYASHI, Hitomi, NAKAGAWA, Masanori
• Format: Journal Article
• Language: Japanese
• Published: The Japan Society of Hepatology 1991
• ISSN: 0451-4203; 1881-3593
• DOI: 10.2957/kanzo.32.169

Human Ito cells (ICs) were isolated from a portion of hepatectomized liver. Various types of cells emerged from a small tissue specimen after 14-20 days of culture. ICs were separated from the cultured cells by triple-layered metrizamide solution gradient centrifugation. The ICs have the main morphological features already reported. Approximately less than 5% of the isolated ICs was positive for vitamin A autofluorescence. However, 2-4 times subcultured ICs, almost devoid of fat-droplets, were able to take up retinol, as the appearance of typical vitamin A fluorescence indicated. Fat-droplets were demonstrated in more than 80% of the subcultured cells. The presence of desmin in the subcultured ICs also demonstrated by immunohistochemical method using polyclonal and monoclonal antidesmin antibodies. The method reported here is capable to harvest approximately 1×106 ICs from 500mg of liver tissue.