NTera 2 cells: a human cell line which displays characteristics expected of a human committed neuronal progenitor cell

J Neurosci Res. 1993 Aug 15;35(6):585-602. doi: 10.1002/jnr.490350603.


We have identified a human cell line with a phenotype resembling committed CNS neuronal precursor cells. NTera 2/cl.D1 (NT2/D1) cells expressed nestin and vimentin, intermediate filament (IF) proteins expressed in neuroepithelial precursor cells, as well as MAP1b, a microtubule-associated protein (MAP) expressed in human neuroepithelium. NT2/D1 cells also expressed the cell adhesion molecules NCAM and N-cadherin which are thought to be important in cell-cell interactions within the neuroepithelium. These NT2/D1 cells also expressed small amounts of NF-L, alpha-internexin, NF-M, and MAP2c, indicating that they are committed to a neuronal fate. Previous studies have shown that, following RA treatment, a proportion of NT2/D1 cells terminally differentiate into neurons and that this occurs via an asymmetric stem cell mode of differentiation. In light of the identification of the neuroepithelial phenotype of NT2/D1 cells we decided to examine more closely the relationship of in vitro neurogenesis in NT2/D1 cells, during RA treatment to that of neurons in vivo. Three days after RA treatment, islands of NT2/D1 cells showed increased expression of neurofilament proteins and increased phosphorylation of NF-M. By 10-14 days, these cells began to resemble neurons morphologically, i.e., with rounded cell bodies and processes. These neuronal cells were clustered into clumps which rested on top of a layer of progenitor cells. In this upper layer, the neurons began to express MAP2b and tau and extinguished their expression of nestin. Recently, we developed a method for obtaining pure cultures of neurons from RA treated NT2/D1 cells. The phenotype of these postmitotic neurons is clearly dissociated from that of the untreated NT2/D1 cells. Given the data obtained in this study and the characterization of the neurons derived from NT2/D1 cells, we propose that NT2/D1 cells are a committed human neuronal precursor cell line which retains some stem cell characteristics and is capable only of terminal differentiation into neurons.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Biomarkers
  • Blotting, Northern
  • Cell Differentiation / physiology
  • Cell Line
  • DNA Probes
  • DNA, Complementary / immunology
  • DNA, Complementary / metabolism
  • Epithelial Cells
  • Fluorescent Antibody Technique
  • Humans
  • Immunohistochemistry
  • Microtubule-Associated Proteins
  • Nerve Tissue Proteins / biosynthesis
  • Neurons / immunology
  • Neurons / metabolism
  • Neurons / physiology*
  • Phenotype
  • Phosphorylation
  • Stem Cells / immunology
  • Stem Cells / metabolism
  • Stem Cells / physiology*


  • Biomarkers
  • DNA Probes
  • DNA, Complementary
  • Microtubule-Associated Proteins
  • Nerve Tissue Proteins
  • microtubule-associated protein 1B