A novel methodology for the establishment of neuroblastoma cell lines from metastatic marrow. Expression of surface markers, neurofilaments, MDR-1 and myc proteins.
A methodology for rapid isolation of neuroblastoma (NB) cells from marrow with metastatic NB cells was developed using a cocktail of five antibodies and magnetic microspheres coated with secondary antibodies. Cells bound to microspheres were released by brief exposure to chymopapain, followed by repeated culture of released cells in serum supplemented DMEM medium and selection for adherent cells. Using this methodology, over 35 primary cell lines were obtained free of contaminating normal cells. Detailed analyses of over 14 cell lines revealed gross differences in cell phenotype, size, morphology development of neurite processes, and doubling time (40h-80 h). All cell lines expressed the 145 kDa neurofilament (NF) and a few expressed the 200 kDa NF, with very little or no expression of the 68 kDa NF. DNA analyses revealed 80% near diploid cell lines. High expression of the MDR-1 protein was detected in 6/22 of the cell lines tested.
Department of Pediatrics Hematology Oncology, University of Florida Medical Center, Gainesville 32610.
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Gazitt
Y J
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Graham-Pole
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000912:Antibodies, Neoplasm; D014408:Biomarkers, Tumor; D016465:Bone Marrow Purging / Q000379:methods; D001859:Bone Neoplasms / Q000378:metabolism* / Q000556:secondary; D002461:Cell Line, Transformed; D004247:DNA / Q000032:analysis; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D006801:Humans; D008562:Membrane Glycoproteins / Q000096:biosynthesis*; D009447:Neuroblastoma / Q000378:metabolism*; D016900:Neurofilament Proteins / Q000096:biosynthesis*; D011003:Ploidies; D016271:Proto-Oncogene Proteins c-myc / Q000096:biosynthesis*
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000912:Antibodies, Neoplasm; D014408:Biomarkers, Tumor; D008562:Membrane Glycoproteins; D016900:Neurofilament Proteins; D016271:Proto-Oncogene Proteins c-myc; D004247:DNA