Human Keratinocytes Synthesize, Secrete, and Deposit Fibronectin in the Pericellular Matrix
Fibronectin production by human keratinocytes cultured in serum-free, low-calcium medium without a fibroblast feeder layer was examined using several techniques. Immunohistochemical examination confirmed that the cultures were not contaminated with fibroblasts or Langerhans cells. By indirect immuno...
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| Published in | Journal of investigative dermatology Vol. 82; no. 6; pp. 580 - 586 |
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| Main Authors | , , , , |
| Format | Journal Article |
| Language | English |
| Published |
Danvers, MA
Elsevier Inc
01.06.1984
Nature Publishing |
| Subjects | |
| Online Access | Get full text |
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| Abstract | Fibronectin production by human keratinocytes cultured in serum-free, low-calcium medium without a fibroblast feeder layer was examined using several techniques. Immunohistochemical examination confirmed that the cultures were not contaminated with fibroblasts or Langerhans cells. By indirect immunofluorescence, fibronectin appeared as intracellular granules within all cells and as short radial fibrils between the cells and the substratum, and in the pericellular matrix. Conditioned media taken from 4-, 6-, 8-, 10-, and 12-day cultures contained fibronectin as measured by enzyme-linked immunosorbent assay in proportion to the cell number in cultures. Synthesis of fibronectin by 7-day keratinocyte cultures was determined by 18-h [35S]methionine metabolic labeling followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis and autoradiography. Fibronectin accounted for 12% of the secreted protein under these culture conditions. Our results indicate that human keratinocytes under conditions that prevent terminal differentiation in vitro can synthesize, secrete, and deposit fibronectin in the extracellular matrix. |
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| AbstractList | Fibronectin production by human keratinocytes cultured in serum-free, low-calcium medium without a fibroblast feeder layer was examined using several techniques. Immunohistochemical examination confirmed that the cultures were not contaminated with fibroblasts or Langerhans cells. By indirect immunofluorescence, fibronectin appeared as intracellular granules within all cells and as short radial fibrils between the cells and the substratum, and in the pericellular matrix. Conditioned media taken from 4-, 6-, 8-, 10-, and 12-day cultures contained fibronectin as measured by enzyme-linked immunosorbent assay in proportion to the cell number in cultures. Synthesis of fibronectin by 7-day keratinocyte cultures was determined by 18-h [35S]methionine metabolic labeling followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis and autoradiography. Fibronectin accounted for 12% of the secreted protein under these culture conditions. Our results indicate that human keratinocytes under conditions that prevent terminal differentiation in vitro can synthesize, secrete, and deposit fibronectin in the extracellular matrix. |
| Author | Howell, Samuel E. Kubo, Miyoko Ryan, Sheli R Norris, David A. Clark, Richard A F |
| Author_xml | – sequence: 1 givenname: Miyoko surname: Kubo fullname: Kubo, Miyoko organization: Department of Dermatology, University of Colorado School of Medicine, Denver, Colorado, U.S.A – sequence: 2 givenname: David A. surname: Norris fullname: Norris, David A. organization: Department of Dermatology, University of Colorado School of Medicine, Denver, Colorado, U.S.A – sequence: 3 givenname: Samuel E. surname: Howell fullname: Howell, Samuel E. organization: Department of Dermatology, University of Colorado School of Medicine, Denver, Colorado, U.S.A – sequence: 4 givenname: Sheli R surname: Ryan fullname: Ryan, Sheli R organization: Department of Dermatology, University of Colorado School of Medicine, Denver, Colorado, U.S.A – sequence: 5 givenname: Richard A F surname: Clark fullname: Clark, Richard A F organization: Department of Dermatology, University of Colorado School of Medicine, Denver, Colorado, U.S.A |
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| Keywords | Human Epidermis Keratinocyte Extracellular matrix Biosynthesis Skin Metabolism Fibronectin |
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57 Boyce (10.1111/1523-1747.ep12261325_bb0250) 1983; 81 Grinnell (10.1111/1523-1747.ep12261325_bb0065) 1979; 17 Nelson (10.1111/1523-1747.ep12261325_bb0150) 1983; 80 Chen (10.1111/1523-1747.ep12261325_bb0010) 1977; 106 Huff (10.1111/1523-1747.ep12261325_bb0195) 1982; 78 Clark (10.1111/1523-1747.ep12261325_bb0120) 1981; 126 Couchman (10.1111/1523-1747.ep12261325_bb0125) 1979; 266 Norris (10.1111/1523-1747.ep12261325_bb0085) 1982; 129 Furie (10.1111/1523-1747.ep12261325_bb0200) 1980; 365 Wigley (10.1111/1523-1747.ep12261325_bb0025) 1979; 118 Laemmli (10.1111/1523-1747.ep12261325_bb0225) 1970; 227 Huang (10.1111/1523-1747.ep12261325_bb0240) 1976; 35 Tsao (10.1111/1523-1747.ep12261325_bb0185) 1982; 110 Lane (10.1111/1523-1747.ep12261325_bb0210) 1982; 79 Wertz (10.1111/1523-1747.ep12261325_bb0270) 1983; 97 Brown (10.1111/1523-1747.ep12261325_bb0165) 1983; 61 Klebe (10.1111/1523-1747.ep12261325_bb0055) 1974; 250 Stanley (10.1111/1523-1747.ep12261325_bb0245) 1983; 96 Sun (10.1111/1523-1747.ep12261325_bb0190) 1978; 14 Clark (10.1111/1523-1747.ep12261325_bb0115) 1982; 79 Rhoads (10.1111/1523-1747.ep12261325_bb0220) 1973; 248 Critchley (10.1111/1523-1747.ep12261325_bb0095) 1979; 280 Rennard (10.1111/1523-1747.ep12261325_bb0205) 1982; II Hennings (10.1111/1523-1747.ep12261325_bb0230) 1980; 19 Gibson (10.1111/1523-1747.ep12261325_bb0130) 1983; 81 Grinnell (10.1111/1523-1747.ep12261325_bb0105) 1981; 76 Alitalo (10.1111/1523-1747.ep12261325_bb0160) 1982; 94 Chandrasekhar (10.1111/1523-1747.ep12261325_bb0255) 1983; 80 Roberts (10.1111/1523-1747.ep12261325_bb0170) 1983; 212 Clark (10.1111/1523-1747.ep12261325_bb0140) 1982; 79 Clark (10.1111/1523-1747.ep12261325_bb0145) 1983; 80 Pearlstein (10.1111/1523-1747.ep12261325_bb0060) 1976; 262 Yamada (10.1111/1523-1747.ep12261325_bb0050) 1978; 275 Stampfer (10.1111/1523-1747.ep12261325_bb0045) 1981; 67 Crouch (10.1111/1523-1747.ep12261325_bb0260) 1978; 78 Mans (10.1111/1523-1747.ep12261325_bb0215) 1961; 94 |
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| SubjectTerms | Biological and medical sciences Enzyme-Linked Immunosorbent Assay Epidermal Cells Epidermis - metabolism Fibronectins - biosynthesis Fibronectins - metabolism Fluorescent Antibody Technique Fundamental and applied biological sciences. Psychology Humans Vertebrates: skin, associated glands, phaneres, light organs, various exocrine glands (salt gland, uropygial gland...), adipose tissue, connective tissue |
| Title | Human Keratinocytes Synthesize, Secrete, and Deposit Fibronectin in the Pericellular Matrix |
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