体外培养小鼠下颌下腺细胞生长特性的实验研究

目的：研究小鼠下颌下腺细胞的培养方法，探讨下颌下腺细胞培养条件及细胞生长特性，为研究干细胞转分化涎腺腺泡细胞以及涎腺再生研究奠定理论基础和技术支持。方法：取2周龄的小鼠，组织块法和消化法分别进行培养，用活细胞观察法和HE染色法记录细胞形态学特征；免疫荧光染色法鉴定；通过生长曲线和细胞倍增时间来比较两种方法对细胞增殖能力的影响。结果：组织块法和消化法均可以成功的培养下颌下腺细胞，（1）组织块法培养的细胞呈卵圆形或多边形，10天左右成铺路石样；消化法培养细胞亦为上皮样细胞，呈多边形，胞质丰富；（2）HE染色下颌下腺细胞呈多边形，胞核明显；（3）Cytokeratin13和AQP5表达阳性，Wimentin表达阴性；（4）组织块法培养细胞增殖相对较平缓，消化法培养细胞增长较迅速；（5）消化法培养倍增时间（1．3471±0．6071）天比组织块法倍增时间（2．1887±1．1503）明显缩短（P〈0．05）；结论：体外可以成功的培养下颌下腺细胞，但是同时证明下颌下腺细胞长期传代较困难，这为研究干细胞转分化涎腺细胞和涎腺再生体内实验提供了理论和实验基础。

The Study of Submandibular Gland Cells Biology Characteristics

Objective： To study the methods of culturing the submandibular gland cells of mouse, and discuss culture conditions and cell growth characteristics of the submandibular gland cells, and aim to laying the theoretical foundation and technical support for the research of stem cells turning to salivary gland acinar cells and salivary gland regeneration. Methods： The 2-week submandibular gland cells were cultured by the tissue block method and the digestion method separately, and then we observed the cell morphology characteristic by living cells photographic and HE dyeing. The cells were identified by immunofluorescence staining method and through the growth curve and cell multiplication time to compare the influence of cell proliferation ability between two methods. Results： Submandibular gland cells can be cultured successfully by organization block method and digestion method. （1） The cells cultured by tissue block method are ovoid or polygon, for 10 days they formed paving-stone sample, digestion culture cell are epithelical cells, polygonal and cytoplasmic rich. （2） HE dyed show the submandibular gland cells are polygonal and the nucleus is obvious. （3） The expression of Cytokeratinl3 and AQP5 are positive and Wimentin is negative. （4） The speed of cell proliferation of tissue block method is relatively gentle, while digestion method is rapid. （5） The doubling time of digestion method （1.3471 ± 0.6071 days ） is significantly shortened than organization block method （2.1887 ± 1.1503） （P〈0.05）. Conclusion： It has been successful to culture submandibular gland cells in vitro, and submandibular gland cells is expected to become seed cells of salivary gland regeneration tissue engineering.