Original Article

Journal of Investigative Dermatology (1995) 105, 557–561; doi:10.1111/1523-1747.ep12323451

Rapid Spreading and Mature Hemidesmosome Formation in HaCaT Keratinocytes Induced by Incubation with Soluble Laminin-5r

Marketta Hormia1,2, Jutta Falk-Marzillier1, George Plopper1, Richard N Tamura1, Jonathan C R Jones3 and Vito Quaranta1

  1. 1Department of Cell Biology, The Scripps Research Institute, La Jolla, California, U.S.A.
  2. 2Department of Periodontology, Institute of Dentistry, University of Helsinki, P.O. Box 41, FIN-00014, Finland
  3. 3Department of Cell, Molecular and Structural Biology, Northwestern University Medical School, Chicago, Illinois, U.S.A.

Received 4 March 1995; Revised 6 June 1995; Accepted 15 June 1995.

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Abstract

HaCaT cells, an immortalized keratinocyte line, incubated in plastic wells in the presence of conditioned medium from 804G cells adhered and spread rapidly in less than 30 min. In contrast, cells plated in fibroblast or keratinocyte conditioned medium adhered poorly and remained rounded at 30 min. Immunodepletion of 804G conditioned medium with polyclonal antisera to laminin-5r, but not control antisera, abolished rapid cell spreading. Electron microscopy of HaCaT cells spread by incubation in 804G conditioned medium, but not control medium, revealed mature hemidesmosomes after 24 h. Rapid spreading was also observed in wells precoated with 804G conditioned medium or 804G cell-deposited matrix, but not with fibronectin, vitronectin, or laminin-1. Immunoblotting of 804G conditioned medium with anti-laminin-5r antibodies unveiled polypeptides of 150, 140, 135, and 100 kDa, identical by electrophoretic mobility to immunoreactive polypeptides in 804G deposited matrix. Our results suggest that addition of laminin-5r in a soluble form is sufficient to promote rapid spreading and hemidesmosome assembly in keratinocytes. The mechanism of soluble laminin-5r action may include efficient surface "priming" for cell adhesion. Soluble laminin-5r may have a physiologic role in morphogenesis and repair of the epidermis and may be of use for therapeutic applications.

Keywords:

cell culture, extracellular matrix, epithelium, wound healing

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