NTera 2 cells: a human cell line which displays characteristics expected of a human committed neuronal progenitor cell.
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.
Pleasure
S J
SJ
Department of Pathology, University of Pennsylvania School of Medicine, Philadelphia.
Lee
V M
VM
D015415:Biomarkers; D015152:Blotting, Northern; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D015342:DNA Probes; D018076:DNA, Complementary / Q000276:immunology / Q000378:metabolism; D004847:Epithelial Cells; D005455:Fluorescent Antibody Technique; D006801:Humans; D007150:Immunohistochemistry; D008869:Microtubule-Associated Proteins; D009419:Nerve Tissue Proteins / Q000096:biosynthesis; D009474:Neurons / Q000276:immunology / Q000378:metabolism / Q000502:physiology*; D010641:Phenotype; D010766:Phosphorylation; D013234:Stem Cells / Q000276:immunology / Q000378:metabolism / Q000502:physiology*
D015415:Biomarkers; D015342:DNA Probes; D018076:DNA, Complementary; D008869:Microtubule-Associated Proteins; D009419:Nerve Tissue Proteins; C085284:microtubule-associated protein 1B