Okura M, Shirasuna K, Hiranuma T, Yoshioka H, Nakahara H, Aikawa T, Matsuya T
First Department of Oral and Maxillofacial Surgery, Faculty of Dentistry, University of Osaka, Japan.
Differentiation. 1993 Sep;54(2):143-53. doi: 10.1111/j.1432-0436.1993.tb00717.x.
Parenchymal tissue of human submandibular glands was cultured in a serum-free medium consisting of a 1:9 mixture of Dulbecco's modified Eagle's medium and MCDB 153 supplemented with 10 ng/ml epidermal growth factor, 10 microM dexamethasone and 1 microgram/ml insulin. Cultivation of the tissue in this medium resulted in propagation of loosely arranged epithelioid cells on plastic, without the necessity of a matrix. Epidermal growth factor significantly enhanced mitogenesis of cultured cells, which expressed specific high- and low-affinity receptors for epidermal growth factor. The epithelioid cells were found to represent the undifferentiated ultrastructure of ductal cells. Immunocytochemically, cultured epithelioid cells expressed antigens specific to basal cells of the intra- and interlobular ducts in situ, including cytokeratins 3 and 6 and cytokeratins 13 and 16, vimentin, and alpha-smooth muscle actin. Moreover, cytoplasm of the cells was immunostained using antibody against the basement membrane component, type IV collagen. These results suggested that cultured epithelioid cells are undifferentiated ductal cells, which have the characteristics of basal cells of the intra- and/or interlobular ducts. Cultured epithelioid cells maintained the characteristics for serial passage until the time that the cultures were confluent. On the other hand, several stratified foci developed on the confluent monolayer. The stratified cells were strongly positive for cytokeratins 3 and 6, but negative for vimentin, alpha-smooth muscle actin and type IV collagen. Moreover, the stratified cells were strongly stained with the antibody against epithelial membrane antigen. This antibody stained the luminal membrane domain of salivary epithelial cells. Electron micrograph of the vertical section through the foci revealed stratified cell layers with a gradual transition from basal cells to squamous epidermoid cells. This result suggests that cultured epithelioid cells, which have the characteristics of basal cells of the intra- and/or interlobular duct, have the potential to differentiate into luminal duct cells.
人下颌下腺实质组织在一种无血清培养基中培养，该培养基由杜氏改良 Eagle 培养基和 MCDB 153 按 1:9 的比例混合而成，并添加了 10 ng/ml 表皮生长因子、10 μM 地塞米松和 1 μg/ml 胰岛素。在这种培养基中培养组织导致在塑料上松散排列的上皮样细胞增殖，无需基质。表皮生长因子显著增强了培养细胞的有丝分裂，这些细胞表达了表皮生长因子的特异性高亲和力和低亲和力受体。发现上皮样细胞代表导管细胞的未分化超微结构。免疫细胞化学分析显示，培养的上皮样细胞原位表达小叶内和小叶间导管基底细胞特有的抗原，包括细胞角蛋白 3 和 6、细胞角蛋白 13 和 16、波形蛋白以及α-平滑肌肌动蛋白。此外，使用针对基底膜成分IV型胶原的抗体对细胞的细胞质进行免疫染色。这些结果表明，培养的上皮样细胞是未分化的导管细胞，具有小叶内和/或小叶间导管基底细胞的特征。培养的上皮样细胞在培养汇合之前保持连续传代的特征。另一方面，汇合的单层上形成了几个分层灶。分层细胞对细胞角蛋白 3 和 6 呈强阳性，但对波形蛋白、α-平滑肌肌动蛋白和IV型胶原呈阴性。此外，分层细胞被抗上皮膜抗原的抗体强烈染色。该抗体可染色唾液上皮细胞的腔面膜区域。通过这些灶的垂直切片的电子显微镜照片显示分层的细胞层，从基底细胞逐渐过渡到鳞状表皮样细胞。这一结果表明，具有小叶内和/或小叶间导管基底细胞特征的培养上皮样细胞具有分化为管腔导管细胞的潜力。
