A simple method for the isolation of human Ito cells (Fat-storing cells)
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 morpho...
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| Published in | Kanzo Vol. 32; no. 2; pp. 169 - 174 |
|---|---|
| Main Authors | , , |
| Format | Journal Article |
| Language | Japanese |
| Published |
The Japan Society of Hepatology
1991
|
| Online Access | Get full text |
| ISSN | 0451-4203 1881-3593 |
| DOI | 10.2957/kanzo.32.169 |
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| Abstract | 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. |
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| AbstractList | 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. |
| Author | NAKAGAWA, Masanori NAKANO, Hiroshi NAKABAYASHI, Hitomi |
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| References | 10) Arenson DM, Friedman SL, Bissell DM: Formation of extracellular matrix in normal rat liver: Lipocytes as a major source of proteoglycan. Gastroenterol 95: 441-447, 1988 17) Davis BH, Pratt BM, Madri JA: Retinol and extracellular collagen matrices modulate hepatic Ito cell collagen phenotype and cellular retinol binding protein levels. J Biol Chem 262: 10280-10286, 1987 18) Miyazaki A, Yokoi Y, Matsuzaki K, et al: Immunological cross-species reactivity of desmin in fat-storing cells (Ito cells) of vertebrates. Acta Histochem Cytochem 19: 219-229, 1986 14) Matsuura T, Nagamori S, Fujise K, et al: Morphological characteristics of human fatstoring cells fractionated and cultured by newly techniques. Human Cell 2: 181-188, 1989 2) Tsutsumi M, Takada A, Takase S, Ooshima A: Connective tissue components in cultured parenchymal and nonparenchymal cells of rat liver; immunohistochemical studies. Lab Invest 58: 88-92, 1988 1) Knook DL, Seffelaar AM, De Leeuw AM: Fat-storing cells of the rat liver, their isolation and purification. Exp Cell Res 139: 468-471, 1982 12) Shiratori Y, Geerts A, Ichida T, et al: Kupffer cells from CCl4-induced fibrotic livers stimulate proliferation of fat-storing cells. J Hepatol 3: 294-303, 1986 7) Kent G, Gay S, Inoue T, et al: Vitamin A containing lipocytes and formation of type III collagen in liver injury. Proc Natl Acad Sci USA 73: 3719-3722, 1976 5) Yokoi Y, Namihisa T, Kuroda H, et al: Immunocytochemical detection of desmin in fatstoring cells (Ito cells). Hepatology 4: 709-714, 1984 8) Friedman SL, Roll FJ, Boyles J, et al: Hepatic lipocyte: The principal collagen-producing cells of normal rat liver. Proc Natl Acad Sci USA 82: 8681-8685, 1985 11) Shiratori Y, Ichida T, Geerts A, et al: Modulation of collagen synthesis by fat-storing cells, isolated from Ca4 or vitamin A treated rats. Digestive Diseases Sciences 32: 1281-1289, 1987 16) 松浦知和,永森静志,藤瀬清隆,他:培養ラットFat-storing Cell (stellate cells)におけるビタミンA移送に関する研究.肝臓 27: 480-486, 1986 3) De Leeuw AM, McCarthy SP, Geerts A, et al: Purified rat liver fat-storing cells in culture divide and contain collagen. Hepatology 4: 392-403, 1984 13) Gressner AM, Zerbe O: Kupffer cell-mediated induction of synthesis and secretion of proteoglycans by rat liver fat-storing cells in culture. J Hepatol 5: 299-310, 1987 6) Tanikawa K: Ultrastructure of hepatic fibrosis and fat-storing cells. In: Collagen Metabolism in the Liver. Editeds by Popper H, Becker K, Stratton Intercontinental Medical Book Co. New York, 1975, p 93-99 9) Clement B, Grimaud JA, Campion JP, et al: Cell types involved in collagen and fibronectin production in normal and fibrotic human liver. Hepatology 6: 225-234, 1986 15) Minato Y, Hasumura Y, Takeuchi J: The role of fat-storing cells in disse space fibrogenesis in alcoholic liver disease. Hepatology 3: 559-566, 1983 4) Chen W, Gendrault JL, Steffan AM, et al: Isolation, culture and main characterstics of mouse fat-storing cells: Interaction with viruses. Hepatology 9: 352-362, 1989 |
| References_xml | – reference: 5) Yokoi Y, Namihisa T, Kuroda H, et al: Immunocytochemical detection of desmin in fatstoring cells (Ito cells). Hepatology 4: 709-714, 1984 – reference: 10) Arenson DM, Friedman SL, Bissell DM: Formation of extracellular matrix in normal rat liver: Lipocytes as a major source of proteoglycan. Gastroenterol 95: 441-447, 1988 – reference: 8) Friedman SL, Roll FJ, Boyles J, et al: Hepatic lipocyte: The principal collagen-producing cells of normal rat liver. Proc Natl Acad Sci USA 82: 8681-8685, 1985 – reference: 9) Clement B, Grimaud JA, Campion JP, et al: Cell types involved in collagen and fibronectin production in normal and fibrotic human liver. Hepatology 6: 225-234, 1986 – reference: 18) Miyazaki A, Yokoi Y, Matsuzaki K, et al: Immunological cross-species reactivity of desmin in fat-storing cells (Ito cells) of vertebrates. Acta Histochem Cytochem 19: 219-229, 1986 – reference: 11) Shiratori Y, Ichida T, Geerts A, et al: Modulation of collagen synthesis by fat-storing cells, isolated from Ca4 or vitamin A treated rats. Digestive Diseases Sciences 32: 1281-1289, 1987 – reference: 16) 松浦知和,永森静志,藤瀬清隆,他:培養ラットFat-storing Cell (stellate cells)におけるビタミンA移送に関する研究.肝臓 27: 480-486, 1986 – reference: 7) Kent G, Gay S, Inoue T, et al: Vitamin A containing lipocytes and formation of type III collagen in liver injury. Proc Natl Acad Sci USA 73: 3719-3722, 1976 – reference: 3) De Leeuw AM, McCarthy SP, Geerts A, et al: Purified rat liver fat-storing cells in culture divide and contain collagen. Hepatology 4: 392-403, 1984 – reference: 1) Knook DL, Seffelaar AM, De Leeuw AM: Fat-storing cells of the rat liver, their isolation and purification. Exp Cell Res 139: 468-471, 1982 – reference: 6) Tanikawa K: Ultrastructure of hepatic fibrosis and fat-storing cells. In: Collagen Metabolism in the Liver. Editeds by Popper H, Becker K, Stratton Intercontinental Medical Book Co. New York, 1975, p 93-99 – reference: 15) Minato Y, Hasumura Y, Takeuchi J: The role of fat-storing cells in disse space fibrogenesis in alcoholic liver disease. Hepatology 3: 559-566, 1983 – reference: 17) Davis BH, Pratt BM, Madri JA: Retinol and extracellular collagen matrices modulate hepatic Ito cell collagen phenotype and cellular retinol binding protein levels. J Biol Chem 262: 10280-10286, 1987 – reference: 4) Chen W, Gendrault JL, Steffan AM, et al: Isolation, culture and main characterstics of mouse fat-storing cells: Interaction with viruses. Hepatology 9: 352-362, 1989 – reference: 2) Tsutsumi M, Takada A, Takase S, Ooshima A: Connective tissue components in cultured parenchymal and nonparenchymal cells of rat liver; immunohistochemical studies. Lab Invest 58: 88-92, 1988 – reference: 12) Shiratori Y, Geerts A, Ichida T, et al: Kupffer cells from CCl4-induced fibrotic livers stimulate proliferation of fat-storing cells. J Hepatol 3: 294-303, 1986 – reference: 13) Gressner AM, Zerbe O: Kupffer cell-mediated induction of synthesis and secretion of proteoglycans by rat liver fat-storing cells in culture. J Hepatol 5: 299-310, 1987 – reference: 14) Matsuura T, Nagamori S, Fujise K, et al: Morphological characteristics of human fatstoring cells fractionated and cultured by newly techniques. Human Cell 2: 181-188, 1989 |
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| Title | A simple method for the isolation of human Ito cells (Fat-storing cells) |
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