Original Article

Journal of Investigative Dermatology (1994) 102, 55–60; doi:10.1111/1523-1747.ep12371732

Epidermis as a Secretory Tissue: An In Vitro Tissue Model to Study Keratinocyte Secretion

Anne B Katz and Lorne B Taichman

Department of Oral Biology and Pathology, School of Dental Medicine, State University of New York at Stony Brook, Stony Brook, New York, U.S.A.

Received 27 May 1993; Accepted 12 June 1993.

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Abstract

In addition to protective functions, keratinocytes secrete a variety of effector molecules that may have local or distant effects. To explore the secretory activity of keratinocytes we have developed a two-chamber culture model in which a fully differentiated stratified epithelium is present in the upper chamber and secreted protein is collected in a lower chamber. A collagen matrix is not used and during collection of secreted protein, 3T3 feeder cells are absent. Keratinocytes secrete protein at the rate of 0.67 mug/h/106 cells, encompassing a molecular weight range from several kD to over 180 kD. Secretion is an unexplored area of keratinocyte biology and this model will allow investigation of this activity.

Keywords:

culture, epithelial, secreted protein

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References

  1. Nozaki, S, Feliciani, C, Sauder, DN: Keratinocyte cytokines. Adv Dermatol 1991 7: 83–101,
  2. Gordon, DA, Fenjves, EF, Williams, DL, Taichman, LB: Synthesis and secretion of apolipoprotein E by cultured human epidermal keratinocytes. J Dermatol 1989 92: 96–99,
  3. Fenjves, ES, Gordon, DA, Penning, LK, Williams, DL, Taichman, LB: Systemic distribution of apolipoprotein E secreted by grafts of epidermal keratinocytes implications for epidermal function and gene therapy. Proc Natl Acad Sci USA 1989 86: 8803–8807,  | PubMed | ChemPort |
  4. Mahley, RW, Apolipoprotein, E: cholesterol transport protein with expanding role in cell biology. Science 1988 240: 622–630,  | PubMed | ISI | ChemPort |
  5. Rheinwald, JG, Green, H: Serial cultivation of strains of human epidermal keratinocytes: the formation of keratinizing colonies from single cells. Cell 1975 6: 331–344,  | Article | PubMed | ISI | ChemPort |
  6. Bell, E, Sher, S, Hull, B: The reconstitution of living skin. J Invest Dermatol 1983 81: 2–10,  | Article | PubMed |
  7. Wu, Y-J, Parker, LM, Binder, NE, Beckett, MA, Sinard, JH, Griffiths, CT, Rheinwald, JG: The mesothelial keratins: a new family of cytoskeletal proteins identified in cultured mesothelial cells and nonkeratinizing epithelia. Cell 1982 31: 693–703,  | Article | PubMed | ISI | ChemPort |
  8. Dunbar, BS, Kimura, H, Timmons, TM: Protein analysis using high-resolution two-dimensional polyacrylamide gel electrophoresis. Meth Enzymol 1990 182: 441–459,
  9. Sambrook, J, Fritsch, EF, Maniatis, T: Molecular Cloning: A Laboratory Manual. 2nd Ed. Cold Spring Harbor Laboratory Press 1989 18.47–18.59,
  10. Bonner, WM, Laskey, RA: A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem 1974 46: 83–88,  | Article | PubMed | ISI | ChemPort |
  11. Sambrook, J, Fritsch, EF, Maniatis, T: Molecular Cloning: A Laboratory Manual, 2nd Ed. Cold Spring Harbor Laboratory Press 1989 18.60–18.75,
  12. Balcarova-Stander, J, Pfeiffer, SE, Fuller, SD, Simons, K: Development of cell surface polarity in the epithelial Madin-Darby canine kidney (MDCK) cell line. EMBO J 1984 3: 2687–2694,  | PubMed | ISI | ChemPort |
  13. Lavker, RM, Matoltsy, AG: Formation of horny cells: the fate of cell organelles and differentiation products in ruminal epithelium. J Cell Biol 1970 44: 501–512,  | Article | PubMed | ChemPort |
  14. Rice, RH, Thatcher, SM: Involucrin: a constituent of cross-linked envelopes and marker of squamous maturation. In: Bereiter-Hahn J, Matoltsy AG, Richards KS (eds.) Biology of the Integument 1986, 2: Springer-Verlag, New York, pp 752–761,
  15. Wu, TL, Soifer, NE, Burtis, WJ, Milstone, LM, Stewart, AF: Glycosylation of parathyroid hormone-related peptide secreted by human epidermal keratinocytes. J Clin Endocrinol Metab 1991 73: 1002–1007,  | PubMed | ISI | ChemPort |
  16. Allan, VJ, Kreis, TE: A microtubule-binding protein associated with membranes of the Golgi apparatus. J Cell Biol 1986 103: 2229–2239,  | Article | PubMed | ISI | ChemPort |
  17. Dover, R, Wright, NA: The cell proliferation kinetics of the epidermis. In: Goldsmith LA (ed.). Physiology, Biochemistry, and Molecular Biology of the Skin, 2nd Edition 1991, Oxford University Press, Oxford, pp 239–265,
  18. Milstone, LM, LaVigne, JF: Heterogeneity of basal keratinocytes: nonrandom distribution of thymidine-labeled basal cells in confluent cultures is not a technical artifact. J Invest Dermatol 1985 84: 504–507,  | Article | PubMed | ISI | ChemPort |
  19. Schermer, A, Jester, JV, Hardy, C, Milano, D, Sun, T-T: Transient synthesis of K6 and K16 keratins in regenerating rabbit corneal epithelium: keratin markers for an alternative pathway of Keratinocyte differentiation. Differentiation 1989 42: 103–110,  | PubMed | ISI | ChemPort |
  20. Elias, PM, McNutt, NS, Friend, DS: Membrane alterations during cornification of mammalian squamous epithelia: a freeze-fracture, tracer and thin-section study. Anat Rec 1977 189: 577–594,  | Article | PubMed | ISI | ChemPort |
  21. Vaughan, FL, Gray, RH, Bernstein, IA: Growth and differentiation of primary rat keratinocytes on synthetic membranes. In Vitro Cell Dev Biol 1986 22: 141–149,
  22. Valyi-Nagy, IT, Murphy, GF, Mancianti, M-L, Whitaker, D, Herlyn, M: Pheno-1 types and infractions of human melanocytes and keratinocytes in an epidermal reconstruction model. Lab Invest 1990 62: 314–324,  | PubMed | ChemPort |
  23. Simons, K, Fuller, SD: Cell surface polarity in epithelia. Ann Rev Cell Biol 1985 1: 243–288,
  24. Matoltsy, AG, Parakkal, PF: Membrane-coating granules of keratinizing epithelia. J Cell Biol 1965 24: 297–307,  | Article | PubMed | ISI | ChemPort |
  25. Clark, RAF, Nielson, LD, Howell, SE, Folkvord, JM: Human keratinocytes that have not terminally differentiated synthesize lamin and fibronectin but deposit only fibroncetin in the pericellular matrix. J Cell Biochem 1985 28: 127–141,
  26. O'Keefe, EJ, Woodley, D, Castillo, G, Russell, N, Payne, RE: Production of soluble and cell-associated fibronectin by cultured keratinocytes. J Invest Dermatol 1984 82: 150–155,
  27. Stanley, JR, Hawley-Nelson, P, Yaar, M, Martin, GR, Katz, SI: Laminin and bullous pemphigoid antigen are distinct basement membrane proteins synthesized by epidermal cells. J Invest Dermatol 1982 78: 456–459,  | Article | PubMed | ISI | ChemPort |
  28. Regauer, S, Seiler, GR, Barrandon, Y, Easley, KW, Compton, CC: Epithelial origin of cutaneous anchoring fibrils. J Cell Biol 1990 111: 2109–2115,  | Article | PubMed | ISI | ChemPort |

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