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J Neurosci Methods. 2008 Sep 15;174(1):31-41. doi: 10.1016/j.jneumeth.2008.06.031. Epub 2008 Jul 10.

Closing the phenotypic gap between transformed neuronal cell lines in culture and untransformed neurons.

Author information

1
Division of Bacteriology & Parasitology, Tulane National Primate Research Center, 18703 Three Rivers Road, Covington, LA 70433, USA.

Abstract

Studies of neuronal dysfunction in the central nervous system (CNS) are frequently limited by the failure of primary neurons to propagate in vitro. Neuronal cell lines can be substituted for primary cells but they often misrepresent normal conditions. We hypothesized that a three-dimensional (3D) cell culture system would drive the phenotype of transformed neurons closer to that of untransformed cells, as has been demonstrated in non-neuronal cell lines. In our studies comparing 3D versus two-dimensional (2D) culture, neuronal SH-SY5Y (SY) cells underwent distinct morphological changes combined with a significant drop in their rate of cell division. Expression of the proto-oncogene N-myc and the RNA-binding protein HuD was decreased in 3D culture as compared to standard 2D conditions. We observed a decline in the anti-apoptotic protein Bcl-2 in 3D culture, coupled with increased expression of the pro-apoptotic proteins Bax and Bak. Moreover, thapsigargin (TG)-induced apoptosis was enhanced in the 3D cells. Microarray analysis demonstrated significantly differing mRNA levels for over 700 genes in the cells of the two culture types, and indicated that alterations in the G1/S cell-cycle progression contributed to the diminished doubling rate in the 3D-cultured SY cells. These results demonstrate that a 3D culture approach narrows the phenotypic gap between neuronal cell lines and primary neurons. The resulting cells may readily be used for in vitro research of neuronal pathogenesis.

PMID:
18672002
PMCID:
PMC2586177
DOI:
10.1016/j.jneumeth.2008.06.031
[Indexed for MEDLINE]
Free PMC Article

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