Functional and Phenotypical Analysis of 19 EuroBoNet Osteosarcoma Cell Lines
AB Mohseny, I Machado, Y Cai, KL Schaefer, PC Hogendoorn, A Llombart-Bosch, AM Cleton-Jansen. Leiden University Medical Center, Leiden, Netherlands; University of Valencia, Valencia, Spain; Shandong University, Jinan, China; University Medical Center Duesseldorf, Duesseldorf, Germany
Background: Recently the EuroBoNet consortium reported the genetic characterization of 36 bone tumour cell lines (DOI: 10.1002/gcc.20717). Here we present the functional analysis of all the osteosarcoma cell lines included there (n=19) regarding their histology, differentiation capacity and tumuorigenicity to identify cell lines that could be considered as representative images of the parental tumours which would be useful as in vitro and in vivo models to study osteosarcoma.
Design: For functional characterization a subset of the cell cultures was pushed towards differentiation into osteoblasts, adipocytes, and chondrocytes using differentiation specific culturing conditions. Furthermore all cell lines were injected subcutaneously and intra muscularly into nude mice to assay their in vivo tumour formation capacity as well as for phenotypical analysis of the tumours. All formed xenografts were further characterized histologically and immunohistochemically using markers involved in differential differentiation (osteonectin, osteocalcin, ALP, cytokeratin, EMA, SMA, desmin and LCA), growth (CD99, BCL2, caveolin1, Ki-67, p53, p16, p21, EGFR, CD34, Her2 and TPD52) and invasion/migration (ezrin, vimentin, e-cadherin and CD31) of osteosarcoma.
Results: 12/16 (75%) cell lines showed adipogenic differentiation of which 8 (8/16, 50%) could also differentiate towards osteoblasts. Only 1/4 (25%) cell lines assayed showed chondrogenic differentiation, interestingly this cell line was not able to differentiate towards adipocytes or osteoblasts. About half of the cell lines (11/19, 58%) produced tumours in vivo after subcutaneous and/or intra muscular injections. All xenografts showed a protein expression pattern representative of high grade osteosarcoma. Both adjacent invasion and lung metastases were observed in some cell lines.
Conclusions: The use of cell lines, especially in cancer research, is of high importance. Questioning their representativeness of clinical tumours should not be addressed by focusing on their genetics exclusively. This study has established a spectrum of osteosarcoma cell lines that robustly represent clinical osteosarcoma. Together with the previously characterized genetic attributes we now have a tool to study the association between genotype and phenotype in a well defined model.
Category: Bone & Soft Tissue
Tuesday, March 23, 2010 9:30 AM
Poster Session III # 14, Tuesday Morning