Application of multicolor fluorescence in situ hybridization analysis for detection of cross-contamination and in vitro progression in commonly used murine tumor cell lines.
Murine tumor models are potent tools for cancer studies, most of which make use of a limited number of murine tumor cell lines that are exchanged by many research groups around the world. Although cross-contamination and in vitro karyotypic progression are well-known risks with respect to the identity of tumor cell lines, these parameters are rarely evaluated. Notably, routine karyotyping of murine cell lines is laborious and technically demanding because mouse chromosomes are morphologically similar. We therefore used a 21-color fluorescence in situ hybridization (FISH) approach (COBRA) for screening two groups of frequently used murine tumor cell lines, each of which shares known immunologic determinants. Multicolor analysis revealed that the sharing of immunologic determinants among three murine lymphoma cell lines (EL-4, MBL-2, and RBL-5) is directly related to their common origin. In several of the cell lines, the chromosomal derivatives had rearranged further, suggesting that the cross-contamination events were not recent. In contrast, karyotypic analysis of three murine colon cancer cell lines (C26, CC36, and C51) showed that these constituted independent tumor clones despite the sharing of immunologic determinants. Our data point out that cross-contamination and in vitro evolution of murine tumor cell lines are a common phenomenon, and that multicolor FISH analysis is an efficient tool for verifying the origin and tracking the evolution of murine cell lines.
Wiegant
Joop
J
Laboratory for Cytochemistry and Cytometry, Department Molecular Cell Biology, Leiden University Medical Center, 2333 AL, Leiden, The Netherlands.
van Hall
Thorbald
T
van der Burg
Marja
M
Colombo
Mario
M
Tanke
Hans J
HJ
Offringa
Rienk
R
Rosenberg
Carla
C
D000230:Adenocarcinoma / Q000235:genetics / Q000473:pathology; D000818:Animals; D000951:Antigens, Neoplasm / Q000235:genetics; D018929:Cell Culture Techniques / Q000379:methods*; D002999:Clone Cells / Q000737:chemistry; D003110:Colonic Neoplasms / Q000235:genetics / Q000473:pathology; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D007621:Karyotyping; D016399:Lymphoma, T-Cell / Q000235:genetics / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D009363:Neoplasm Proteins / Q000235:genetics; D013945:Thymoma / Q000235:genetics / Q000473:pathology; D013953:Thymus Neoplasms / Q000235:genetics / Q000473:pathology; D014407:Tumor Cells, Cultured / Q000737:chemistry* / Q000276:immunology
D000951:Antigens, Neoplasm; D009363:Neoplasm Proteins