Establishment of human colon cancer cell lines from fresh tumors versus xenografts: Comparison of success rate and cell line features
Establishment of human colon cancer cell lines from fresh tumors versus xenografts: Comparison of success rate and cell line features
复制标题
DOI：
10.1158/0008-5472.can-06-0594
复制
发表时间：
2007-01-01
期刊：
影响因子：
11.2
通讯作者：
Poupon, Marie-France
中科院分区：
文献类型：
--
作者：
Dangles-Marie, Virginie;Pocard, Marc;Poupon, Marie-France
从新鲜肿瘤中获得代表性的人结肠癌细胞系在技术上是困难的。使用来自结肠癌患者的32个肿瘤片段，本研究显示，与从新鲜肿瘤直接建立相比，预先异种移植导致更有效的细胞系建立（P < 0.05）。从26个肿瘤标本中，我们成功地在裸鼠中建立了20个肿瘤异种移植物（77%）;在这些异种移植物中的19个中，9个（47%）导致细胞系，包括4个来自肝转移。31例肿瘤标本中仅3例（9.7%）在体外立即生长，且均来自原发肿瘤。为了比较使用两种方案衍生自相同肿瘤片段的人结肠癌细胞系的主要表型和基因型特征，广泛研究了从32个肿瘤片段中的2个获得的两对细胞系。它们表现出相似的形态，并能够形成紧凑的球体。对5-氟尿嘧啶、CPT 11和L-OHP的化学敏感性在从患者肿瘤获得的细胞系和从异种移植物获得的细胞系之间不同。匹配的细胞系有一个共同的核心核型改变和独特的额外的染色体畸变。通过实时定量PCR评价的肿瘤发生中的作用，发现选择的基因的表达水平在统计学上相关，无论使用的体外培养模型。总之，在建立细胞系之前将肿瘤碎片异种移植到小鼠体内，能够产生更多新型人类癌细胞系，用于研究结肠癌细胞生物学，为复制这种疾病的多样性提供了机会。
Obtaining representative human colon cancer cell lines from fresh tumors is technically difficult. Using 32 tumor fragments from patients with colon cancer, the present study shows that prior xenograft leads to more efficient cell line establishment compared with direct establishment from fresh tumors (P < 0.05). From 26 tumor specimens, we successfully established 20 tumor xenografts in nude mice (77%); among 19 of these xenografts, 9 (47%) led to cell lines, including four from liver metastases. Only 3 of 31 tumor specimens (9.7%) grew immediately in vitro, and all were derived from primary tumors. To compare major phenotypic and genotypic characteristics of human colon cancer cell lines derived from the same tumor fragment using two protocols, the two pairs of cell lines obtained from 2 of 32 tumor fragments were extensively studied. They displayed similar morphology and were able to form compact spheroids. Chemosensitivity to 5-fluorouracil, CPT11, and L-OHP differed between cell lines obtained from patient tumors and those derived from xenografts. Matched cell lines shared a common core of karyotype alterations and distinctive additional chromosomal aberrations. Expression levels of genes selected for their role in oncogenesis evaluated by real-time quantitative PCR were found to be statistically correlated whatever the in vitro culture model used. In conclusion, xenotransplantation in mice of tumor fragments before establishment of cell lines enables generation of more novel human cancer cell lines for investigation of colon cancer cell biology, opening up the opportunity of reproducing the diversity of this disease.