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Establishment of Human Colon Cancer Cell Lines from Fresh Tumors versus Xenografts: Comparison of Success Rate and Cell Line Features

¹ Physiopathologie Hépatique, UMR 8149 Centre National de la Recherche Scientifique, Faculté des Sciences Pharmaceutiques et Biologiques de Paris, Université René Descartes-Paris; ² Instabilité des Microsatellites et Cancers, U762 Institut National de la Sante et de la Recherche Medicale, Université Pierre et Marie Curie-Paris; ³ Pharmacologie Expérimentale, U612 Institut National de la Sante et de la Recherche Medicale, Institut Curie; ⁴ Département d'Anatomie Pathologique, Institut Mutualiste Montsouris; ⁵ Origine, Structure et Evolution de la Biodiversité, UMR 5202 Centre National de la Recherche Scientifique, Museum National d'Histoire Naturelle, Paris, France; ⁶ Département de Chirurgie and ⁷ Département de Biologie Clinique, Institut Gustave-Roussy, Villejuif, France; and ⁸ Service de Médecine Nucléaire, Centre René Huguenin, Saint Cloud, France

Requests for reprints: Virginie Dangles-Marie, Laboratoire de Physiopathologie Hépatique, UMR 8149 Centre National de la Recherche Scientifique, Faculté des Sciences Pharmaceutiques et Biologiques de Paris, Université René Descartes-Paris 5, 4 Avenue de l'Observatoire, 75006 Paris, France. Phone: 33-1-43-29-76-08; Fax: 33-1-43-29-76-12; E-mail: virginie.dangles{at}univ-paris5.fr.

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. [Cancer Res 2007;67(1):398–407]

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