基础医学与临床 ›› 2017, Vol. 37 ›› Issue (7): 1010-1014.

• 研究论文 • 上一篇    下一篇

人腹膜假黏液瘤原代细胞培养和鉴定

张海红1,张保华1,王亚南2   

  1. 1. 中国医学科学院基础医学研究所
    2. 中国医学科学院基础医学研究所&北京协和医学院基础医学院
  • 收稿日期:2017-03-30 修回日期:2017-05-16 出版日期:2017-07-05 发布日期:2017-06-23
  • 通讯作者: 王亚南 E-mail:shlwangyanan@gmail.com
  • 基金资助:
    中国医学科学院医学与健康科技创新工程

Culture and characterization of human pseudomyxoma peritonei cells

  • Received:2017-03-30 Revised:2017-05-16 Online:2017-07-05 Published:2017-06-23

摘要: 目的 体外培养和扩增腹膜假黏液瘤原代细胞,为腹膜假黏液瘤的研究及药物筛选提供基础。方法 用胶原酶消化法分离5例腹膜假黏液瘤样本并进行原代细胞培养,然后用染色体核型分析鉴定培养的细胞是否为肿瘤细胞;再用PAS染色检测细胞中性黏多糖;3D细胞培养模拟肿瘤在体内的状态;HE染色观察3D细胞的组织形态;裸鼠成瘤实验检测原代细胞的致瘤性。结果 腹膜假黏液瘤细胞原代培养5例均成功;2D培养的细胞贴壁增殖,形态为多边形鹅卵石样排列,可传代到18代;染色体核型分析显示大多数细胞是亚二倍体核型的肿瘤细胞;细胞PAS染色阳性,说明培养的原代细胞内含有黏液;3D培养的类器官与体内肿瘤组织形态相似;裸鼠成瘤率低,且形成的肿瘤与患者PMP相似度不高。结论 腹膜假黏液瘤原代细胞可在体外培养和扩增。

关键词: 关键词:腹膜假粘液瘤, 原代培养, 3D培养

Abstract: Objective Culture, expansion and characterization of pseudomyxoma peritonei cells, which laid the foundation for the study of pseudomyxoma peritonei and drug screening. Methods Tumors from 5 cases of human pseudomyxoma peritonei were cultured by collagenase digestion. The cultured cells were then identified as tumor cells by chromosome karyotyping. The secretion of neutral mucopolysaccharide was detected by PAS staining. To simulate the tumor growth condition in vivo, 3D culture was applied and the morphology of the cultured organoid was observed with HE staining, xenografts were used to test the tumorigenicity of PMP primary cells. Results The primary culture of pseudomyxoma peritonei cells was successful in all of the 5 cases. 2D cultured cells were adherent, polygonal pebble-like arranged, and passed on up to 18 generations. Chromosome karyotype analysis showed that most of the cells were subdiploid karyotype tumor cells. Positive PAS staining suggests that these cells secrete mucus. The morphology of 3D cultured organoid was similar to that of tumor tissues. The tumor formation rate of xenografts was low, and the tumoris not similar to patient’s PMP. Conclusion Pseudomyxoma peritonei tumor cells can be cultured and expanded in vitro.

Key words: Key words:Pseudomyxoma peritonei, Primary culture , 3D culture

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