人甲状腺乳头状癌原代细胞的体外长期培养  

Long-Term Primary Culture of Human Papillary Thyroid Carcinoma Cells

作　　者：边学海[1] 赵吉生[2] 张秀英[3] 张纯海[1] 付言涛[1] 赵丽娜[1] 孙辉[1] 郑泽霖[2] 

机构地区：[1]吉林大学中日联谊医院甲状腺外科,吉林长春130033 [2]吉林大学中日联谊医院胃肠外科,吉林长春130033 [3]吉林大学护理医学院,吉林长春130031

出　　处：《中国普外基础与临床杂志》2011年第8期840-843,共4页Chinese Journal of Bases and Clinics In General Surgery

摘　　要：目的探索人甲状腺乳头状癌原代细胞的体外长期培养方法及其监测鉴定手段。方法常规分离甲状腺乳头状癌细胞,并用含10%胎牛血清、L-谷氨酰胺和20 ng/ml人重组表皮生长因子（EGF）的高糖型DMEM培养液培养,分别在不同时间观察甲状腺乳头状癌细胞生长情况,测定培养液中的甲状腺球蛋白（Tg）和甲状腺过氧化物酶（TPO）水平,用免疫细胞化学方法检测Tg的表达。结果培养约8 d甲状腺乳头状癌细胞长满约80%的培养瓶底,符合传代标准;在培养45 d内生长良好,经5-6传代后细胞逐渐老化。细胞培养液中Tg含量随培养时间延长呈抛物线形变化,培养约14 d Tg达到峰值（985.2μg/L）;TPO在细胞培养液中始终未检测到。免疫荧光染色结果显示,体外培养甲状腺乳头状癌原代细胞的Tg表达阳性。结论应用本研究建立的培养条件可以使人甲状腺乳头状癌原代细胞传5-6代,细胞存活约45 d。建立了简便、系统的监测及鉴定甲状腺乳头状癌细胞生物活性手段,提示在14 d左右该细胞用于基因研究较为合适,为研究甲状腺乳头状癌发生、发展机理及其特征提供了一种合适替代工具。Objective To investigate the primary culture method of human papillary thyroid carcinoma（PTC） cells for a long term and establish a monitoring and verification measures.Methods PTC cells were isolated following routine procedures and cultured in the DMEM supplemented with 10% fetal bovine serum,glutamine,and 20 ng/ml epidermal growth factor（EGF）.Thyroglobulin（Tg） and thyroperoxidase（TPO） in nutrient solution and specific antigen Tg expression of PTC cells cultured for different days were observed.Results The PTC cells grew satisfactorily up to 45 days of incubation.Tg content in nutrient solution expressed the training period of a linear singular parabolic,achieved peak value（985.2 μg/L） at about 14 d.TPO had not been detected in nutrient solution.The Tg expressed positively by immunization fluorescent dyeing.Conclusions PTC cells cultured in the present method can survive to over 45 days.A brief monitoring and evaluation systems of PTC cells has been established.This report prompts that cultured cells within 14 days maybe more suitable to gene research and provide alternative to the basic research of PTC events and features.

关 键 词：甲状腺乳头状癌 原代细胞培养 甲状腺球蛋白 甲状腺过氧化物酶 免疫组织化学 

分 类 号：R736.1[医药卫生—肿瘤]