人胃癌原代细胞短期培养与纯化培养体外药敏试验  

Chemosensitivity testing for human gastric cancer with short-term culture cells vs purified primary culture cells using the MTT assay

作　　者：许洪伟[1] 秦成勇[1] 朱菊人[1] 刘吉勇[1] 王春霞[2] 劳萍[2] 张春清[1] 

机构地区：[1]山东省立医院消化内科,山东济南250021 [2]山东省立医院中心实验室,山东济南250021

出　　处：《中国癌症杂志》2003年第3期243-246,共4页China Oncology

基　　金：山东省医药卫生科研基金 (编号 2 0 0 1 CA2DBB2 )

摘　　要：目的 :评价人胃癌原代细胞短期培养 (混杂非肿瘤细胞 )与纯化培养MTT法体外药敏试验的可行性、各自的优势与缺陷。方法 :采用短期培养MTT比色法测定 7种化疗药物敏感性。另经反复贴壁、胶原酶消化排除、胰蛋白酶消化选择或自然纯化法获得纯的原代肿瘤细胞 ,再行MTT法。测定两种方法细胞数与A(光密度 )值的相关性、药物抑制率及两种方法药敏结果的灵敏性。结果 :短期法MTT药敏试验成功率 81 4% (48/59) ;纯化培养为 50 8% (30 /59) ;两种方法细胞数与A值均呈直线相关 (P均 <0 0 1 ) ,药敏结果具有可比性 ;两种方法 7种药物敏感性顺序一致 ,但纯化法显示 7种化疗药物的抑制率有 6种均高于短期法 ,表明纯化法较敏感 ;纯化培养所需时间平均 (2 0 2± 9 5)天。结论 :人胃癌原代细胞短期培养MTT比色法快捷、简便 ,能够反映化疗药敏结果的基本趋势及个体化差异 ,但灵敏性略低。纯化培养后药敏试验结果更理想 。Purpose:To evaluate the feasibility, advantages and disadvantages of chemosensitivity test of human gastric cancer using MTT assay with short term culture tumor cells contaminated by nonmalignant cells compared with purifiedly primary culture tumor cells.Methods:Fifty nine fresh samples from patients with gastric cancer were obtained from operating rooms. Chemosensitivity results were provided by the MTT assay with short term culture cells. The primary culture cells were purified by means of a series of methods such as repeated cell attachment, differential trypsinization and natural purification which removed fibroblasts and other nonmalignant cells. The same MTT assay was conducted using purified cells. Chemosensitivity results between short term method and purification method were compared for seven antitumor drugs. Results:The success rate of short term method using the MTT assay for chemosensitivity testing was 81 4% (48 of 59 patients),and for the purification method it was 50.8(30 of 59 patients). The average was 20.2±9.5 days when primary cells were cultured into purified cancer cells. The optical density ( A ) value correlated directly with the the number of tumor cells with the two methods( P <0.01). All of the seven antitumor drugs but one using purified cells showed higher inhibition rates than those using short term method.Conclusions:Chemosensitivity testing of human gastric cancer with short term culture cells using the MTT assay was a rapid, simple and reliable method. Its degree of accuracy was slightly poorer, than purified primary cell culture. Chemosensitivity with purified primary culture cells was a sensitive method at the expense of longer interval, but then its success rate was not satisfactory. [

关 键 词：胃癌 原代细胞 培养 纯化 体外 药敏试验 

分 类 号：R735.2[医药卫生—肿瘤]