Establishment and characterization of a human neuroblastoma cell line

A continuous human cell line RN-GA was established from a stage-III primary neuroblastoma prior to therapy. Light and electron microscopic analysis of the biopsy showed morphological features typical of neuroectodermal origin. Relative cellular DNA content and N-myc oncogene copy number were also analyzed in the biopsy tissue: the tumor cells presented a near-diploid genome with N-myc amplification. The derived tumor cell line expressed distinctive ultrastructural, cytogenetic and immunological markers of neuroblastoma. Moreover, cells from the culture could be serially transplanted into splenectomized-irradiated nude mice, where they formed a progressively growing solid tumor. Surprisingly, the cells in culture did not show any N-myc amplification, while they retained a near-diploid DNA content. We propose that several techniques (electron microscopy, oncogene analysis, flow cytometry, cytogenetics, tissue culture, cell antigen immunodetection) should be used to establish a firm diagnosis and a correct clinical grading of this tumor. The establishment of this continuous cell line should be valuable as an experimental in vitro system for further studies of neuroblastoma biology and morphology.