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Characterization of astrocytes derived from human NTera-2/D1 embryonal carcinoma cells.
J Neurosci Res. 2002 Jun 01; 68(5):604-14.JN

Abstract

Astrocytes are the predominant cell type in the vicinity of glutamatergic synapses, where they monitor and maintain low levels of glutamate. Synaptic homeostasis of glutamate involves its removal from the synaptic cleft via high-affinity glutamate transporters, glutamate transporter-1 (GLT-1)/excitatory amino acid transporters (EAAT)2 and glutamate and aspartate transporter (GLAST)/EAAT1, and glutamate-catabolizing enzyme, glutamine synthase. Glutamate transporters have been mostly characterized in rodent astrocytes, due to the lack of a convenient human cell system. We report here that NTera-2 (NT2/D1, a cell line derived from a human teratocarcinoma and known to differentiate into neurons) can also be differentiated by a 4-week treatment with retinoic acid into functional astrocytes (NT2/A). Differentiation was accompanied by decreased cell proliferation and cell-cycle arrest, as measured by flow cytometry, immunostaining for Ki67 and incorporation of 5-bromo-2'deoxyuridine (BrdU). Immunocytochemistry and Western blot analysis showed that NT2/A expressed glial fibrillary acidic protein, vimentin and S100beta. Reverse transcription polymerase chain reaction (PCR) detected mRNA encoding glutamate transporters GLT-1/EAAT2 and GLAST/EAAT1. The expression level of GLAST/EAAT1 was higher than that of GLT-1/EAAT2, which is a typical expression pattern for primary astrocytes. Functionality of the transporters was demonstrated by the uptake of (3)H-glutamate. NT2/A also expressed active glutamine synthase, and treatment with glutamate (up to 1 mM for 24 hr) was non-toxic, suggesting that these cells were capable of converting it to non-toxic metabolites. NT2/A and NT2-derived neurons could be grown as mixed cultures and this may prove to be a useful experimental model to study molecular mechanisms underlying glutamate excitotoxicity.

Authors+Show Affiliations

Apoptosis Research Group, Neurobiology Program, Institute for Biological Sciences, National Research Council of Canada, Ottawa, Ontario, Canada. jagdeep.sandhu@nrc.caNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

12111850

Citation

Sandhu, Jagdeep K., et al. "Characterization of Astrocytes Derived From Human NTera-2/D1 Embryonal Carcinoma Cells." Journal of Neuroscience Research, vol. 68, no. 5, 2002, pp. 604-14.
Sandhu JK, Sikorska M, Walker PR. Characterization of astrocytes derived from human NTera-2/D1 embryonal carcinoma cells. J Neurosci Res. 2002;68(5):604-14.
Sandhu, J. K., Sikorska, M., & Walker, P. R. (2002). Characterization of astrocytes derived from human NTera-2/D1 embryonal carcinoma cells. Journal of Neuroscience Research, 68(5), 604-14.
Sandhu JK, Sikorska M, Walker PR. Characterization of Astrocytes Derived From Human NTera-2/D1 Embryonal Carcinoma Cells. J Neurosci Res. 2002 Jun 1;68(5):604-14. PubMed PMID: 12111850.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Characterization of astrocytes derived from human NTera-2/D1 embryonal carcinoma cells. AU - Sandhu,Jagdeep K, AU - Sikorska,Marianna, AU - Walker,P Roy, PY - 2002/7/12/pubmed PY - 2002/8/13/medline PY - 2002/7/12/entrez SP - 604 EP - 14 JF - Journal of neuroscience research JO - J. Neurosci. Res. VL - 68 IS - 5 N2 - Astrocytes are the predominant cell type in the vicinity of glutamatergic synapses, where they monitor and maintain low levels of glutamate. Synaptic homeostasis of glutamate involves its removal from the synaptic cleft via high-affinity glutamate transporters, glutamate transporter-1 (GLT-1)/excitatory amino acid transporters (EAAT)2 and glutamate and aspartate transporter (GLAST)/EAAT1, and glutamate-catabolizing enzyme, glutamine synthase. Glutamate transporters have been mostly characterized in rodent astrocytes, due to the lack of a convenient human cell system. We report here that NTera-2 (NT2/D1, a cell line derived from a human teratocarcinoma and known to differentiate into neurons) can also be differentiated by a 4-week treatment with retinoic acid into functional astrocytes (NT2/A). Differentiation was accompanied by decreased cell proliferation and cell-cycle arrest, as measured by flow cytometry, immunostaining for Ki67 and incorporation of 5-bromo-2'deoxyuridine (BrdU). Immunocytochemistry and Western blot analysis showed that NT2/A expressed glial fibrillary acidic protein, vimentin and S100beta. Reverse transcription polymerase chain reaction (PCR) detected mRNA encoding glutamate transporters GLT-1/EAAT2 and GLAST/EAAT1. The expression level of GLAST/EAAT1 was higher than that of GLT-1/EAAT2, which is a typical expression pattern for primary astrocytes. Functionality of the transporters was demonstrated by the uptake of (3)H-glutamate. NT2/A also expressed active glutamine synthase, and treatment with glutamate (up to 1 mM for 24 hr) was non-toxic, suggesting that these cells were capable of converting it to non-toxic metabolites. NT2/A and NT2-derived neurons could be grown as mixed cultures and this may prove to be a useful experimental model to study molecular mechanisms underlying glutamate excitotoxicity. SN - 0360-4012 UR - https://www.unboundmedicine.com/medline/citation/12111850/Characterization_of_astrocytes_derived_from_human_NTera_2/D1_embryonal_carcinoma_cells_ L2 - https://doi.org/10.1002/jnr.10236 DB - PRIME DP - Unbound Medicine ER -