title: A basement membrane-like matrix formed by cell-released proteins at the medium/air interface supports growth of keratinocytes.
abstract: Epithelial cells require adherence to a matrix for regular growth. During standard keratinocyte cell culture in serum-free medium, we observed that cell colonies formed not only on the bottom of the culture vessels but also at the medium/air interface. Coomassie blue staining detected a protein membrane that extended up to several centimeters between the colonies of floating cells. Ultrastructural investigation of this membrane revealed structures closely resembling those of basement membranes, and immunochemical staining confirmed the presence of laminins-1 and -5 as well as collagen IV, representative components of basement membranes. Cells attached to the floating membrane proliferated and could be cultivated for up to six months. When keratinocyte-conditioned medium was filtered and transferred to a culture vessel without cells, the protein membrane at the liquid/air interface formed within one week suggesting self-assembly of cell-released proteins. Our findings provide a basis for the production of epidermal basement membranes for potential medical uses.
authors.affiliation: Department of Dermatology, University of Vienna Medical School, Vienna, Austria.
authors.forename: Leopold
authors.initials: L
authors.lastname: Eckhart
authors.forename: Christina
authors.initials: C
authors.lastname: Reinisch
authors.forename: Sadayuki
authors.initials: S
authors.lastname: Inoue
authors.forename: Paul
authors.initials: P
authors.lastname: Messner
authors.forename: Michael
authors.initials: M
authors.lastname: Dockal
authors.forename: Christoph
authors.initials: C
authors.lastname: Mayer
authors.forename: Erwin
authors.initials: E
authors.lastname: Tschachler
mesh_terms: D001485:Basement Membrane / Q000166:cytology* / Q000378:metabolism; D002448:Cell Adhesion / Q000502:physiology; D002455:Cell Division / Q000502:physiology; D002478:Cells, Cultured; D024141:Collagen Type IV / Q000378:metabolism*; D016895:Culture Media, Serum-Free / Q000737:chemistry; D000078404:Epidermal Cells; D004817:Epidermis / Q000378:metabolism; D006801:Humans; D015603:Keratinocytes / Q000166:cytology* / Q000378:metabolism; D007797:Laminin / Q000378:metabolism*; D008854:Microscopy, Electron
chemical_list: D024141:Collagen Type IV; D016895:Culture Media, Serum-Free; D007797:Laminin