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

• Published in: Journal of neuroscience research Vol. 35; no. 6; p. 585
• Main Authors: Pleasure, S J, Lee, V M
• Format: Journal Article
• Language: English
• Published: United States 15.08.1993
• Subjects: 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
• ISSN: 0360-4012
• 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.