体外培养大鼠颌下腺细胞的生物学特性研究

Biological characteristics of cultured rat submandibular gland cells in vitro

目的 :研究体外培养颌下腺细胞生物学特性。方法 :应用胰酶消化法进行颌下腺细胞的分离纯化 ,然后进行颌下腺细胞原代和传代培养。绘制培养细胞的生长曲线 ;MTT法检测培养细胞活力大小。透射电镜 (TEM )观察结合免疫组化检测鉴定细胞来源及性质。结果 :整个颌下腺细胞生长期间为呈多样性的上皮细胞 ,细胞可传 4～ 5代 ,成活 3 0～ 40d。随培养时间延长 ,各组细胞数量都有不同程度的增加 ,比较培养 72h原代、第 2代与第 4代细胞吸光度值 ,原代、第 2代与第 4代相比较有显著性差异 (P <0 .0 5 )。超微结构观察及免疫组化染色培养细胞表现为腺上皮细胞的特征。结论 :原代和传代培养第 1代、第 2代颌下腺细胞活性较强 。

Objective:To investigate the biological characteristics of cultured salivary gland cells in vitro. Methods: Submandibular gland cells of rat were primarily cultured and subcultured in DMEM supplemented with φ=10% fetal bovine serum. The growth of the cells was studied by cell counting and MTT assay. The morphological characteristic was observed with microscope and transmission electron microscope.The specific protein expression was investigated by immunohistochemical technic. Results:Morpholgy of the cells was epidermoid with well developed organella and zymogen granules.The cells could subultured to 4～5 passages and kept in culture for 30～40 days.The cells of primary culture and passage 2 grew faster than those of passage 4.The cells were CK8.13, keratin and α-SMA positive. Conclusion:Rat submandibular gland cells can be primarily cultured and subcultured in vitro.The primarily cultured and first subcultured cells may be more potential in growth.