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Differentiation of the human NT2 cells into neurons and glia

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Methods in Cell Science

Abstract

A method underlying a strategy for differentiation of the NT2 human teratocarcinoma cell line into neuronal and glial cells is described. The aim of this work is to provide a human model to study the relationships between neurons and glia in vitro during developmental or degenerative events. NT2 cells are seeded on polylysine precoated plastic or glass and differentiated by all-trans retinoic acid; persistent undifferentiated cells are eliminated by cytosine-β-D-arabinofuranoside; then cell cultures are maintained during four weeks until the appearance of glutamatergic receptors. Along the differentiation procedure, we have followed the expression of neuronal and glial phenotypes as well as the excitotoxic response to N-methyl-D-aspartate treatment taken as an indication of neuronal maturation. The procedure described leads to the development of a mixed population of neurons and glia sensitive to glutamate exposure.

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References

  1. Andrews PW (1984). Retinoic acid induces neuronal differentiation of a cloned human embryonal carcinoma cell line in vitro. Dev Biol 103: 285–293. 0

    PubMed  Google Scholar 

  2. Andrews PW, Damjanov I, Simon D, Banting GS, Carlin C, Dracopoli NC, Fogh J (1984). Pluripotent embryonal carcinoma clones derived from the human teratocarcinoma cell line Tera-2. Lab Invest 50(2): 147–162. 0

    PubMed  Google Scholar 

  3. Andrews PW (1988). Human teratocarcinomas. Biochim Biophys Acta 948: 17–36. 0

    PubMed  Google Scholar 

  4. Attwell D (1994). Glia and neurons in dialogue. Nature 369: 707–708. 0

    Google Scholar 

  5. Barres BA (1991). New roles for glia. J Neurosci 11: 3685–3694. 0

    PubMed  Google Scholar 

  6. Cheung WMW, Chu AH, IP NY (1997). Identification of candidate genes induced by retinoic acid in embryonal carcinoma cells. J Neurochem 68(5): 1882–1888. 0

    PubMed  Google Scholar 

  7. Fogh J, Trempe G (1975). Human tumor cells in vitro. In: Fogh J (ed), New human tumor cell lines, pp 115–159. New York: Plenum. 0

    Google Scholar 

  8. Hardy M, Younkin D, Tang C-M, Pleasure J, Shi QY, Williams M, Pleasure D (1994). Expression of non-NMDA glutamate receptor channel genes by clonal human neurons. J Neurochem 63(2): 482–489. 0

    PubMed  Google Scholar 

  9. Jones-Villeneuve EM, Rudnicki MA, Harris JF, McBurney MW (1983). Retinoic acid-induced neural differentiation of embryonal carcinoma cells. Mol Cell Biol 3: 2271–2279.

    PubMed  Google Scholar 

  10. Koh JY, Choi DW (1987). Quantitative determination of glutamate mediated cortical neuronal injury in cell culture by lactate dehydrogenase efflux assay. J Neurosci Meth 20: 83–90.

    Google Scholar 

  11. Kurie JM, Buch J, Eppinger TM, Moy D, Dmitrovsky E (1993). 9-cis and all-trans retinoic acid induce a similar phenotype in human teratocarcinoma cells. Differentiation 54: 123–129.

    PubMed  Google Scholar 

  12. Kurie JM, Younes A, Miller WH, Burchert M, Chiu C-F, Kolesnich R, Dmitrovsky E (1993). Retinoic acid stimulates the protein kinase C pathway before of its H5E-nucleor receptor during human teratocarcinoma differentiation. Biochim Biophys Acta 1179: 203–207.

    PubMed  Google Scholar 

  13. Kurie JM, Allopenna J, Dmitrovsky E (1994). Retinoic acid stimulates protein kinase A-associated G proteins during human teratocarcinoma differentiation. Biochim Biophys Acta 1222: 88–94.

    PubMed  Google Scholar 

  14. Miller WH, Moy D, Li A, Grippo JF, Dmitrovsky E (1990) Retinoic acid induces down-regulation of several growth factors and proto-oncogenes in a human embryonal cancer cell line. Oncogene 5: 511–517.

    PubMed  Google Scholar 

  15. Moasser MM, DeBlasio A, Dmitrovsky E (1994). Response and resistance to retinoic acid are mediated through the retinoic acid nuclear receptor γ in human teratocarcinomas. Oncogene 9: 833–840.

    PubMed  Google Scholar 

  16. Munir M, Lu L, McGonigle P (1995). Excitotoxic cell death and delayed rescue in human neurons derived from NT2 cells. J Neurosci 15(12): 7847–7860.

    PubMed  Google Scholar 

  17. Munir M, Lu L, Wang Y-H, Luo J, Wolfe BB, McGonigle P (1996). Pharmacological and immunological characterization on N-methyl-D-aspartate receptors in human NT2-N neurons. J Pharmacol Exp Ther 276(2): 819–828.

    PubMed  Google Scholar 

  18. Parpura V, Basarky TA, Liu F, Jeftinija K, Jeftinija S, Haydon PG (1994). Glutamate-mediated astrocyteneuron signalling. Nature 369: 744–747.

    PubMed  Google Scholar 

  19. Pfrieger FW, Barres BA (1996). New views on synapse-glia interactions. Curr Opin Neurobiol 6: 615–621.

    PubMed  Google Scholar 

  20. Pleasure SJ, Page C, Lee VM-Y (1992). Pure, post-mitotic, polarized human neurons derived from Ntera 2 cells provide a system for expressing exogenous proteins in terminally differentiated neurons. J Neurosci 12(5): 1802–1815.

    PubMed  Google Scholar 

  21. Pleasure SJ, Lee VM-Y (1993). Ntera 2 cells: A human cell line which displays characteristics expected of a human committed neuronal progenitor cell. J Neurosci Res 35: 585–602.

    PubMed  Google Scholar 

  22. Rudnicki MA, McBurney MW (1987). Teratocarcinomas and embryonic stem cells. In: Robertson J (ed), Cell culture methods and induction of differentiation of embryonal carcinoma cell lines, pp 19–49. Oxford: IRL Press.

    Google Scholar 

  23. Staines WA, Morassutti DJ, Reul KR, Ally AI, McBurney MW (1994). Neurons derived from embryonal carcinoma cells have varied morphologies and neurotransmitters. Neurosciences 58: 735–751.

    Google Scholar 

  24. Stemple DL, Anderson DJ (1992). Isolation of a stem cell for neurons and glia from the mammalian neural crest. Cell 71: 973–985.

    Article  PubMed  Google Scholar 

  25. Temple S, Xueming Q (1996). Vertebrate neural progenitor cells: Subtypes and regulation. Curr Opin Neurobiol 6: 11–17.

    PubMed  Google Scholar 

  26. Wolf BA, Wertkin AM, Jolly YC, Yasuda RP, Wolfe BB, Konrad RJ, Manning D, Ravi S, Williamson JR, Lee M-Y (1959). Muscarinic regulation of Alzheimer's disease amyloid precursor protein secretion and amyloid β-protein production in human neuronal NT2N cells. J Biol Chem 270(9): 4916–4922.

    Google Scholar 

  27. Wroblewski F, La Due JS (1955). Lactic dehydrogenase activity in blood. Proc Soc Exp Biol Med 90: 210–213.

    PubMed  Google Scholar 

  28. Younkin DP, Tang C-M, Hardy M, Reddy UR, Shi QY, Pleasure SJ, Lee V-Y, Pleasure D (1995). Inducible expression of neuronal glutamate receptor channels in the NT2 human cell line. Proc Natl Acad Sci USA 90: 2174–2178.

    Google Scholar 

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Langlois, A., Duval, D. Differentiation of the human NT2 cells into neurons and glia. Methods Cell Sci 19, 213–219 (1997). https://doi.org/10.1023/A:1009731707443

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