兔腮腺腺细胞的体外培养和功能评价  

Culture of rabbit parotid acinar cells and evaluation of its excretion function in vitro

作　　者：何巍[1] 乔永明[1] 李龙江[2] 

机构地区：[1]郑州大学第一附属医院口腔颌面外科,河南郑州450052 [2]四川大学口腔生物医学工程教育部重点实验室,四川成都610041

出　　处：《上海口腔医学》2008年第2期165-169,共5页Shanghai Journal of Stomatology

基　　金：国家教委博士点基金(20040610079)

摘　　要：目的:体外培养获取成活率高且保持功能的兔腮腺腺细胞。方法:采用改良DMEM培养液进行兔腮腺腺细胞的原代及传代培养,观察腮腺腺细胞的生长状况;免疫细胞化学法检测细胞中泛角蛋白及α-淀粉酶蛋白和肌动蛋白的表达情况以鉴定细胞来源;检测α-淀粉酶蛋白在不同代次腺细胞中的表达情况,判断其分泌功能。应用SPSS10.0软件包对数据进行t检验。结果:体外培养的兔腮腺腺细胞具有一定的增殖能力,可传3代,存活4周以上;其特异性抗体泛角蛋白及α-淀粉酶蛋白染色为阳性,肌动蛋白染色阴性;不同代次腺细胞α-淀粉酶的表达强度随传代次数的增加而逐渐减弱,原代培养和传代培养第1代的细胞,α-淀粉酶的表达强度较高且可维持一定水平,两者无显著差异(P<0.05);传代培养第2代的细胞,α-淀粉酶含量明显下降,与前者差异显著(P<0.05);第3代细胞基本测不到α-淀粉酶的表达。结论:采用改良DMEM培养液体外培养兔腮腺腺细胞,可得到足量的腺细胞,且前2代细胞保持了较强的分泌功能,是选作实验研究的最佳阶段。PURPOSE： To obtain a large number of rabbit parotid acinar cells with excretion function in vitro. METHODS： Rabbit parotid acinar cells were cultured in the reformed DMEM; The shape and structure of cultured cells were observed with phase contrast microscope; Pan cytokeratin （PCK）, α-amylase and actin of the cultured parotid acinar cells were examined with immunocytochemical SP technique; the secretion of different generations was evaluated with positive expression status of α-amylase. SPSS10.0 software package for windows was used for Student＇s t test. RESULTS： The acinar cells had the ability of proliferation, they could pass 4 generations and survive for 4 weeks. The specific immunocytochemical staining for PCK and α-amylase could be seen in the plasm and （or） membrane of parotid acinar cells,.but not for actin. The positive expression of α-amylase in the primary generation, generationl,2 and 3 was 79.6%, 76% ,47.7% and 2.4%, respectively, which decreased gradually. There was a significant difference between the second generation and two earlier generations（P〈0.05 ）. CONCLUSION： The primary and first generations of parotid acinar cells maintain their characteristics and proliferation in vitro,which is very useful for further experiment. Supported by Doctorate Foundation of Ministry of Education of China （Grant No.20040610079）.

关 键 词：细胞培养 腮腺腺细胞 Α-淀粉酶 兔 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]