PMID- 18672002
OWN - NLM
STAT- MEDLINE
DCOM- 20081106
LR  - 20211203
IS  - 0165-0270 (Print)
IS  - 0165-0270 (Linking)
VI  - 174
IP  - 1
DP  - 2008 Sep 15
TI  - Closing the phenotypic gap between transformed neuronal cell lines in culture and 
      untransformed neurons.
PG  - 31-41
LID - 10.1016/j.jneumeth.2008.06.031 [doi]
AB  - 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.
FAU - Myers, Tereance A
AU  - Myers TA
AD  - Division of Bacteriology & Parasitology, Tulane National Primate Research Center, 
      18703 Three Rivers Road, Covington, LA 70433, USA.
FAU - Nickerson, Cheryl A
AU  - Nickerson CA
FAU - Kaushal, Deepak
AU  - Kaushal D
FAU - Ott, C Mark
AU  - Ott CM
FAU - Honer zu Bentrup, Kerstin
AU  - Honer zu Bentrup K
FAU - Ramamurthy, Rajee
AU  - Ramamurthy R
FAU - Nelman-Gonzalez, Mayra
AU  - Nelman-Gonzalez M
FAU - Pierson, Duane L
AU  - Pierson DL
FAU - Philipp, Mario T
AU  - Philipp MT
LA  - eng
GR  - P51 RR000164-46/RR/NCRR NIH HHS/United States
GR  - RR 00164/RR/NCRR NIH HHS/United States
GR  - R01 NS048952-04/NS/NINDS NIH HHS/United States
GR  - P51 RR000164/RR/NCRR NIH HHS/United States
GR  - R01 NS048952/NS/NINDS NIH HHS/United States
GR  - NS 048952/NS/NINDS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20080710
PL  - Netherlands
TA  - J Neurosci Methods
JT  - Journal of neuroscience methods
JID - 7905558
RN  - 0 (Apoptosis Regulatory Proteins)
RN  - 0 (Cell Cycle Proteins)
RN  - 0 (ELAV Proteins)
RN  - 0 (ELAV-Like Protein 4)
RN  - 0 (ELAVL4 protein, human)
RN  - 0 (Intracellular Signaling Peptides and Proteins)
RN  - 0 (MAS1 protein, human)
RN  - 0 (NMI protein, human)
RN  - 0 (Proto-Oncogene Mas)
RN  - 0 (Proto-Oncogene Proteins c-myc)
RN  - 0 (RNA, Messenger)
SB  - IM
MH  - Animals
MH  - Apoptosis/genetics
MH  - Apoptosis Regulatory Proteins/genetics
MH  - Cell Culture Techniques/methods
MH  - Cell Cycle Proteins/genetics
MH  - Cell Differentiation/physiology
MH  - Cell Division/physiology
MH  - Cell Line, Transformed
MH  - Cell Proliferation
MH  - Cell Shape/physiology
MH  - ELAV Proteins/genetics
MH  - ELAV-Like Protein 4
MH  - Gene Expression Profiling
MH  - Genes, cdc/physiology
MH  - Humans
MH  - Intracellular Signaling Peptides and Proteins
MH  - Neurons/*cytology/*metabolism
MH  - Oligonucleotide Array Sequence Analysis
MH  - Organ Culture Techniques/methods
MH  - PC12 Cells
MH  - Phenotype
MH  - Proto-Oncogene Mas
MH  - Proto-Oncogene Proteins c-myc/genetics
MH  - RNA, Messenger/analysis/genetics
MH  - Rats
PMC - PMC2586177
MID - NIHMS69271
EDAT- 2008/08/02 09:00
MHDA- 2008/11/07 09:00
PMCR- 2009/09/15
CRDT- 2008/08/02 09:00
PHST- 2008/04/22 00:00 [received]
PHST- 2008/06/18 00:00 [revised]
PHST- 2008/06/19 00:00 [accepted]
PHST- 2008/08/02 09:00 [pubmed]
PHST- 2008/11/07 09:00 [medline]
PHST- 2008/08/02 09:00 [entrez]
PHST- 2009/09/15 00:00 [pmc-release]
AID - S0165-0270(08)00380-4 [pii]
AID - 10.1016/j.jneumeth.2008.06.031 [doi]
PST - ppublish
SO  - J Neurosci Methods. 2008 Sep 15;174(1):31-41. doi: 
      10.1016/j.jneumeth.2008.06.031. Epub 2008 Jul 10.