Morphological and immunocytochemical characterization of cultured rat incisor cervical epithelial cells.
Epithelial cells from the cervical loop of the rat incisor were isolated by co-culture of apical explants with growth-arrested 3T3 fibroblasts. The epithelial phenotype of the expanding outgrowths was confirmed 10 days after the seeding of the explants by phase-contrast microscopy and immunocytochemical identification of cytokeratins. After 3 weeks in culture, the epithelial cells covered the entire surface of the coverslips and were then passaged. Subcultures gave rise to a confluent sheet within 10-12 days. Light and electron microscopy showed that confluent cervical epithelial cells generally reconstituted a bi-layered structure similar to Hertwig's epithelial sheath. Epithelial cells from the rat palate, cultured and subcultured according to the same procedure, organized themselves in 5-6 cell layers, the upper cells having generally a squamous morphology. Synthesis of extracellular matrix molecules by rat incisor cervical epithelial cells was studied with specific antibodies. These cells failed to produce type I collagen, but synthesized all the major basement membrane components (type IV collagen, laminin, heparan sulphate proteoglycan and fibronectin). These observations suggest that the culture conditions allowed the reconstitution of a typical Hertwig's epithelial sheath by rat incisor cervical epithelial cells.
Farges
J C
JC
Laboratoire d'Histophysiologie et de Pathologie des Tissus Dentaires, Faculté d'Odontologie, CNRS UPR 412, Lyon, France.
Couble
M L
ML
Joffre
A
A
Hartmann
D J
DJ
Magloire
H
H
D000818:Animals; D002478:Cells, Cultured / Q000166:cytology / Q000648:ultrastructure; D003470:Culture Media / Q000592:standards; D003804:Dentin / Q000166:cytology / Q000254:growth & development* / Q000648:ultrastructure; D004847:Epithelial Cells; D004848:Epithelium / Q000254:growth & development / Q000648:ultrastructure; D005069:Evaluation Studies as Topic; D007150:Immunohistochemistry; D007180:Incisor; D010641:Phenotype; D051381:Rats
D003470:Culture Media