基础医学与临床 ›› 2017, Vol. 37 ›› Issue (2): 224-229.

• 研究论文 • 上一篇    下一篇

乳腺癌组织细胞的高效原代培养及其特性

杨振丽1,徐雅丽1,卞晓翠2,冯海凉3,刘玉琴2,孙强4   

  1. 1. 中国医学科学院基础医学研究所 北京协和医学院基础学院
    2. 中国医学科学院基础医学研究所;北京协和医学院基础学院
    3. 中国协和医科大学基础医学院,中国医学科学院基础所
    4. 北京协和医学院
  • 收稿日期:2016-10-25 修回日期:2016-12-19 出版日期:2017-02-05 发布日期:2017-01-16
  • 通讯作者: 刘玉琴 E-mail:ccc@pumc.edu.cn

Faciliated primary culture and amplification of breast cancer cells and their biological properties

  • Received:2016-10-25 Revised:2016-12-19 Online:2017-02-05 Published:2017-01-16
  • Contact: Yu-qin LIU, E-mail:ccc@pumc.edu.cn

摘要: 目的 高效扩增乳腺癌组织细胞,明确其特性,为乳腺癌深入研究及个体化治疗提供实验材料。方法 利用小鼠胚胎成纤维细胞3T3swiss作饲养层细胞,培养基中添加ROCK抑制剂Y-27632,对乳腺癌组织细胞进行原代培养。 CCK-8检测细胞增殖;流式细胞术检测细胞周期;免疫细胞化学检测细胞CK分子表达;RT-PCR检测HER-2,ER,PR及乳腺癌干细胞相关标志分子(如CD44,CD24等)mRNA的表达;STR检测以明确细胞身份。结果 在饲养层细胞和Y-27632抑制剂同时存在的情况下,所培养的乳腺癌组织细胞增殖速度较快,可短时间获得大量细胞;细胞呈多角形上皮样,CK、HER-2、ER表达阳性,PR 不表达; CD44和CD24表达阳性;经STR分析证明其身份唯一。结论:饲养层细胞及ROCK抑制剂有助于乳腺癌组织细胞的原代培养及大量扩增,扩增的细胞性质稳定,可用于个体化治疗研究。

关键词: 3T3Swiss, 饲养层细胞, Y-27632, 原代培养, 乳腺癌

Abstract: Objective To efficiently builds up and expand breast cancer cells from patient cancer tissue and identify their biological properties, to provide abundant materials for research and personalized medicine. Methods Feeder cell layer and ROCK inhibitor Y-27632 were employed to faciliate the breast cancer cells;CCK-8 was used to determine the proliferation of the breast cancer cells; Cell cycle distribution was analyzed by flow cytometry; Histochemistry(FH) assay to show the expression level of CK. The mRNA expression levels of HER-2, ER, PR and the breast cancer stem cell associated molecules (such as CD44, CD24, etc.) were detected by RT-PCR;STR assay was used for identifying verification of the cells. Results The use of feeder cells and Y-27632 facilitates rapid expand of the original breast cancer cells, and the cells have kept the original features of the tumor. Conclusion To use the method could obtain a large number of cells within a short time, which can promptly be used for the research of personalized medicine.

Key words: 3T3Swiss, feeder cells, Y-27632, primary culture, breast cancer