Farges J C, Couble M L, Joffre A, Hartmann D J, Magloire H
Laboratoire d'Histophysiologie et de Pathologie des Tissus Dentaires, Faculté d'Odontologie, CNRS UPR 412, Lyon, France.
Arch Oral Biol. 1991;36(10):737-45. doi: 10.1016/0003-9969(91)90040-2.
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.
通过将切牙颈部环的顶端外植体与生长停滞的3T3成纤维细胞共培养，分离出大鼠切牙颈部环的上皮细胞。在接种外植体10天后，通过相差显微镜和细胞角蛋白的免疫细胞化学鉴定，证实了生长的上皮细胞表型。培养3周后，上皮细胞覆盖了盖玻片的整个表面，然后进行传代培养。传代培养物在10-12天内形成汇合的细胞层。光镜和电镜显示，汇合的颈部上皮细胞通常重建了类似于赫特维希上皮鞘的双层结构。按照相同程序培养和传代培养的大鼠腭部上皮细胞形成5-6层细胞，上层细胞通常具有鳞状形态。用特异性抗体研究了大鼠切牙颈部上皮细胞的细胞外基质分子合成。这些细胞不能产生I型胶原，但能合成所有主要的基底膜成分（IV型胶原、层粘连蛋白、硫酸乙酰肝素蛋白聚糖和纤连蛋白）。这些观察结果表明，培养条件允许大鼠切牙颈部上皮细胞重建典型的赫特维希上皮鞘。
