title: Comprehensive characterization of neuroblastoma cell line subtypes reveals bilineage potential similar to neural crest stem cells.
abstract: BACKGROUND
  Neuroblastic tumors (NBT) derive from neural crest stem cells (NCSC). Histologically, NBT are composed by neuroblasts and Schwannian cells. In culture, neuroblastic (N-), substrate-adherent (S-) and intermediate phenotype (I-) cell subtypes arise spontaneously.
  
  
  METHODS
  Here, neuroblastoma (NB) cell line subtypes were characterized according to embryonic peripheral nervous system development markers (GAP43, Phox2b, Sox10, c-kit, GD2, NF68, vimentin, S100beta, calcyclin and ABCG2), morphological features, gene expression and differentiation potential. I-type cells were investigated as a bipotential (neuronal and glial) differentiation stage.
  
  
  RESULTS
  Positive immunostaining of NCSC (GAP43, c-kit, NF68, vimentin and Phox2b) and undifferentiated cell (ABCG2) markers was observed in all NB subtypes. N- and I-type cells displayed cytoplasmic membrane GD2 staining, while nuclear calcyclin was restricted to S-type. N- and I-type cells showed similar phenotype and immunoreactivity pattern. Differential gene expression was associated with each cell subtype. N- and I-type cells displayed similar differentiation capacity towards neuronal and glial lineage fates. S-type cells, upon induction, did not show a neuronal-like phenotype, despite gene expression changes.
  
  
  CONCLUSIONS
  Results suggest that N- and I-type NB cell subtypes represent an immature bilineage stage, able to progress towards neuronal and glial fates upon induction of differentiation. S-type cells appear irreversibly committed to a glial lineage fate.
authors.affiliation: Developmental tumor biology laboratory, Hospital Sant Joan de Déu, Barcelona, Spain. sacosta@fsjd.org
authors.forename: Sandra
authors.initials: S
authors.lastname: Acosta
authors.forename: Cinzia
authors.initials: C
authors.lastname: Lavarino
authors.forename: Raquel
authors.initials: R
authors.lastname: Paris
authors.forename: Idoia
authors.initials: I
authors.lastname: Garcia
authors.forename: Carmen
authors.initials: C
authors.lastname: de Torres
authors.forename: Eva
authors.initials: E
authors.lastname: Rodríguez
authors.forename: Helena
authors.initials: H
authors.lastname: Beleta
authors.forename: Jaume
authors.initials: J
authors.lastname: Mora
mesh_terms: D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters / Q000378:metabolism; D001973:Bromodeoxyuridine / Q000494:pharmacology; D018797:Cell Cycle Proteins / Q000235:genetics; D002454:Cell Differentiation / Q000187:drug effects; D045744:Cell Line, Tumor; D019070:Cell Lineage / Q000235:genetics / Q000502:physiology*; D019922:GAP-43 Protein / Q000378:metabolism; D018398:Homeodomain Proteins / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D017350:N-Acetylgalactosaminyltransferases / Q000235:genetics; D009363:Neoplasm Proteins / Q000378:metabolism; D009432:Neural Crest / Q000166:cytology*; D009447:Neuroblastoma / Q000378:metabolism* / Q000473:pathology*; D016900:Neurofilament Proteins / Q000378:metabolism; D019009:Proto-Oncogene Proteins c-kit / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000075383:S100 Calcium Binding Protein A6; D009418:S100 Proteins / Q000235:genetics; D055757:SOXE Transcription Factors / Q000235:genetics; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D014212:Tretinoin / Q000494:pharmacology; D014746:Vimentin / Q000378:metabolism
chemical_list: C116614:ABCG2 protein, human; D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters; D018797:Cell Cycle Proteins; D019922:GAP-43 Protein; D018398:Homeodomain Proteins; C109220:NBPhox protein; D009363:Neoplasm Proteins; D016900:Neurofilament Proteins; D000075383:S100 Calcium Binding Protein A6; D009418:S100 Proteins; C527923:SOX10 protein, human; D055757:SOXE Transcription Factors; D014157:Transcription Factors; D014746:Vimentin; C044882:neurofilament protein L; C055351:S100A6 protein, human; D014212:Tretinoin; D017350:N-Acetylgalactosaminyltransferases; C023220:(N-acetylneuraminyl)-galactosylglucosylceramide N-acetylgalactosaminyltransferase; D019009:Proto-Oncogene Proteins c-kit; D001973:Bromodeoxyuridine