原代人宫颈癌细胞的培养方法  

Culture methods for primary cervical carcinoma cells

作　　者：程静新[1,2] 马秀萍 周萍[1] 袁敏[1] 黄坤[1] 张怡[2] 

机构地区：[1]新疆医科大学附属肿瘤医院妇科,新疆维吾尔自治区乌鲁木齐市831100 [2]中南大学湘雅医院妇科,湖南省长沙市410008 [3]昌吉市第二人民医院,新疆维吾尔自治区昌吉市831100

出　　处：《中国组织工程研究》2013年第15期2769-2775,共7页Chinese Journal of Tissue Engineering Research

基　　金：新疆维吾尔自治区自然科学基金资助项目(2010211A31);课题名称:宫颈癌个体化新辅助化疗模式探讨~~

摘　　要：背景:在体外分离、培养宫颈癌上皮细胞,从细胞水平及分子水平研究肿瘤病因及治疗的基础,是宫颈癌组织工程研究的最基本环节。目的:探讨适用于组织工程宫颈癌研究的人宫颈癌原代细胞培养方法以及传代后细胞形态变化、增殖的特性。方法:0.25%胰蛋白酶和2%Ⅰ型胶原酶联合消化法体外培养宫颈癌原代细胞,荧光倒置显微镜下观察肿瘤细胞形态、体外生长情况,免疫组化法检测体外培养宫颈癌细胞表面标记物CK17和肿瘤细胞增殖抗原Ki67表达。结果与结论:采用胰蛋白酶与Ⅰ型胶原酶联合消化法体外分离培养人宫颈癌细胞,该法相对简单且重复性较好,所得到的原代细胞纯度较高。经体外传代培养的宫颈癌细胞仍可保持稳定的细胞表型。宫颈癌细胞表面标记物CK17阳性表达,证实细胞为上皮源性,肿瘤细胞增殖抗原Ki67表达阳性提示所培养细胞具有肿瘤细胞恶性增殖的特性。BACKGROUND： In vitro isolation and culture of cervical epithelial carcinoma cells is the foundation for studying tumor etiology and treatment from the cellular level, which is the most basic aspect of cervical cancer tissue engineering research. OBJECTIVE： To investigate the culture methods for primary cervical carcinoma cells, and passaged celgrowth, proliferation and shape in vitro. METHODS： We isolated and cultured primary cervical carcinoma cell by using enzyme joint digestion technique （0.25% trypsin and 2% type Ⅰ collagenase）, observed the shape of the tumor cell and condition of the cell proliferation using the inverted fluorescent microscope, and detected CK17 and Ki67 expression by immunofluorescence. RESULTS AND CONCLUSION： Cervical epithelial carcinoma cells harvested using the enzyme joint digestion technique had a higher purity, and this technique was relatively simple to be repeated. The passaged cervical epithelial carcinoma cells also had a stable cell phenotype. CK17 positive expression indicated the cultured cells were cervical epithelial carcinoma cells and Ki67 positive expression showed that the cultured cells had the proliferative characteristics of malignant tumor cells.

关 键 词：组织构建 组织构建细胞学实验 细胞培养 宫颈 上皮细胞 培养基 胰蛋白酶 Ⅰ型胶原酶 原代细胞 体外培养 生长特点 肿瘤标记物 CK17 KI67 省级基金 

分 类 号：R318[医药卫生—生物医学工程]