A novel methodology for the establishment of neuroblastoma cell lines from metastatic marrow. Expression of surface markers, neurofilaments, MDR-1 and myc proteins

Yair Gazitt, Yun Ju He, John Graham-Pole

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)171-178
Number of pages8
JournalJournal of Immunological Methods
Volume148
Issue number1-2
DOIs
StatePublished - 1992
Externally publishedYes

Keywords

  • Bone marrow metastasis
  • MDR-1
  • Neuroblastoma
  • Oncogene

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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