Abstract
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.
Highlights
Colorectal cancer accounts for 10% to 15% of all cancers and is the second leading cause of cancer-related death in industrialized countries
Tumor material not required for histopathologic diagnosis was both snap-frozen in liquid nitrogen and placed in ‘‘collecting medium’’ (DMEM supplemented with 10 mmol/L HEPES, 4.5 g/L glucose, 1 mmol/L pyruvate sodium, 200 units/mL penicillin, 200 Ag/mL streptomycin, 200 Ag/mL gentamicin, 5 Ag/mL ciprofloxacine, 20 Ag/mL metronidazole, 25 Ag/mL vancomycin, and 2.5 Ag/mL fungizone) for further establishment of cell lines or xenografts as follows
Thirty-two colon cancers originated from primary tumors and/or metastases were included in this cell line establishment study
Summary
Colorectal cancer accounts for 10% to 15% of all cancers and is the second leading cause of cancer-related death in industrialized countries. Two major genetic pathways, characterized by microsatellite and chromosomal instability, have been identified in the development of colon cancer [7, 8]. In addition to this disease-related heterogeneity, intrinsic tumor heterogeneity is observed, which is a landmark of colon cancer. Gain and loss of chromosomes in the malignant cell population is considered a process of diversification that leads to survival of the fittest clones [9, 10]. Extensive genotypic and phenotypic heterogeneity is recognized as a major trait of cancer cell populations, this heterogeneity could be rapidly masked by the expansion of a limited number of clones
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