title: Basement membrane formation by malignant mouse keratinocyte cell lines in organotypic culture and transplants: correlation with degree of morphologic differentiation.
abstract: Six malignant C3H mouse epidermal cell lines (HEL-30, HEL-37, HELP I, HELP IV, HD II, H3L), with different capacities for epidermal differentiation, were analyzed for their organized growth behavior and basement membrane (BM) formation in organotypical cultures in vitro and after transplantation into syngeneic mice. Expression and deposition of five BM components (type IV collagen, laminin, bullous pemphigoid antigen, fibronectin, heparan sulfate proteoglycan) were determined on frozen sections by indirect immunofluorescence. Additionally, synthesis and secretion of BM components by the line HEL-30 (in submersed cultures) were identified by metabolic labeling and immunoprecipitation. Morphologic differentiation features and formation of a structured BM were studied by electron microscopy. All cell lines were tumorigenic and invasive but nevertheless able to synthesize BM constituents in vitro and in vivo, although pronounced variations in the expression and the polarity and continuity of deposition were found. Irrespective of the amount of BM components synthesized, none of the cell lines formed a structured BM in organotypical cultures in vitro. After transplantation the production of BM components was improved and the newly formed epithelia were separated from the mesenchyme by a structured BM. The formation of BM occurred whether the epithelial cells were in immediate contact with the mesenchyme or separated by a 1 to 2 mm thick native collagen gel. Deposition of BM constituents and formation of BM structures occurred both at the superficial epithelium and around invading cell cords. The studies clearly demonstrated that malignant epidermal cells maintain their capacity to synthesize BM components. The extent of production and the polarity of deposition of the constituents and the quality of BM formation were usually higher in well differentiated cell lines and obviously correlated well with their preserved degree of differentiation. Comparable to normal keratinocytes, formation of structured BM requires interaction with living mesenchyme but occurs independently of direct epidermal-mesenchymal contact.
mesh_terms: D000818:Animals; D001485:Basement Membrane / Q000648:ultrastructure; D002460:Cell Line; D002471:Cell Transformation, Neoplastic / Q000473:pathology* / Q000648:ultrastructure; D003094:Collagen / Q000032:analysis; D004848:Epithelium / Q000473:pathology / Q000648:ultrastructure; D008565:Membrane Proteins / Q000032:analysis; D051379:Mice; D008809:Mice, Inbred C3H; D008854:Microscopy, Electron; D009368:Neoplasm Transplantation; D009924:Organ Culture Techniques; D012867:Skin / Q000473:pathology* / Q000648:ultrastructure; D014185:Transplantation, Isogeneic
chemical_list: D008565:Membrane Proteins; D003094:Collagen