title: Optimising parameters for the differentiation of SH-SY5Y cells to study cell adhesion and cell migration.
abstract: BACKGROUND
  Cell migration is a fundamental biological process and has an important role in the developing brain by regulating a highly specific pattern of connections between nerve cells. Cell migration is required for axonal guidance and neurite outgrowth and involves a series of highly co-ordinated and overlapping signalling pathways. The non-receptor tyrosine kinase, Focal Adhesion Kinase (FAK) has an essential role in development and is the most highly expressed kinase in the developing CNS. FAK activity is essential for neuronal cell adhesion and migration.
  
  
  RESULTS
  The objective of this study was to optimise a protocol for the differentiation of the neuroblastoma cell line, SH-SY5Y. We determined the optimal extracellular matrix proteins and growth factor combinations required for the optimal differentiation of SH-SY5Y cells into neuronal-like cells and determined those conditions that induce the expression of FAK. It was confirmed that the cells were morphologically and biochemically differentiated when compared to undifferentiated cells. This is in direct contrast to commonly used differentiation methods that induce morphological differentiation but not biochemical differentiation.
  
  
  CONCLUSIONS
  We conclude that we have optimised a protocol for the differentiation of SH-SY5Y cells that results in a cell population that is both morphologically and biochemically distinct from undifferentiated SH-SY5Y cells and has a distinct adhesion and spreading pattern and display extensive neurite outgrowth. This protocol will provide a neuronal model system for studying FAK activity during cell adhesion and migration events.
authors.affiliation: Department of Life Sciences and Materials and Surface Science Institute, University of Limerick, Limerick, Ireland. Patrick.Kiely@ul.ie.
authors.forename: Susan
authors.initials: S
authors.lastname: Dwane
authors.forename: Edel
authors.initials: E
authors.lastname: Durack
authors.forename: Patrick A
authors.initials: PA
authors.lastname: Kiely
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects; D018929:Cell Culture Techniques / Q000592:standards*; D002454:Cell Differentiation / Q000187:drug effects; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D016326:Extracellular Matrix Proteins / Q000494:pharmacology; D051417:Focal Adhesion Kinase 1 / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D009414:Nerve Growth Factors / Q000494:pharmacology; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D015398:Signal Transduction
chemical_list: D015415:Biomarkers; D016326:Extracellular Matrix Proteins; D009414:Nerve Growth Factors; D051417:Focal Adhesion Kinase 1; C496050:PTK2 protein, human
references.citation: Cell Motil Cytoskeleton. 2003 Jun;55(2):77-96
references.pmid: 12740870
references.citation: Annu Rev Neurosci. 1999;22:511-39
references.pmid: 10202547
references.citation: Annu Rev Cell Biol. 1987;3:443-92
references.pmid: 3318882
references.citation: Annu Rev Biochem. 1979;48:193-216
references.pmid: 382984
references.citation: J Biol Chem. 2002 Jul 12;277(28):25297-304
references.pmid: 12000752
references.citation: Mol Cell Endocrinol. 1985 Mar;39(3):169-87
references.pmid: 2984061
references.citation: Cell Motil Cytoskeleton. 1996;35(3):188-99
references.pmid: 8913640
references.citation: Int J Biochem Cell Biol. 1997 Mar;29(3):401-14
references.pmid: 9202420
references.citation: Int J Cancer. 1981 Nov 15;28(5):583-9
references.pmid: 7309295
references.citation: Curr Biol. 2010 Jun 22;20(12):1086-92
references.pmid: 20493699
references.citation: Biosci Rep. 1987 Sep;7(9):681-99
references.pmid: 3322422
references.citation: Neurotox Res. 2004;5(8):579-98
references.pmid: 15111235
references.citation: J Neurosci Res. 2009 Oct;87(13):2821-32
references.pmid: 19437543
references.citation: Proc Natl Acad Sci U S A. 1984 Apr;81(8):2562-6
references.pmid: 6326132
references.citation: Cold Spring Harb Perspect Biol. 2010 Feb;2(2):a001834
references.pmid: 20182622
references.citation: Trends Cell Biol. 2004 Jan;14(1):1-5
references.pmid: 14729173
references.citation: Lab Invest. 1978 Aug;39(2):77-89
references.pmid: 682602
references.citation: Cell. 1986 Jul 18;46(2):155-69
references.pmid: 3013421
references.citation: Brain Res. 2006 Jan 19;1069(1):31-8
references.pmid: 16414032
references.citation: Chin Med J (Engl). 2010 Apr 20;123(8):1086-92
references.pmid: 20497720
references.citation: Front Biosci. 2003 May 01;8:s676-82
references.pmid: 12700130
references.citation: Annu Rev Neurosci. 2003;26:441-83
references.pmid: 12626695
references.citation: Cell Differ. 1983 Mar;12(3):165-70
references.pmid: 6299586
references.citation: Neurotoxicology. 2009 Jan;30(1):127-35
references.pmid: 19056420
references.citation: J Biol Chem. 2003 Jan 3;278(1):391-9
references.pmid: 12401808
references.citation: Nat Neurosci. 2004 Nov;7(11):1213-21
references.pmid: 15494734
references.citation: Cell Differ. 1984 Jun;14(2):135-44
references.pmid: 6467378
references.citation: Matrix Biol. 1997 Oct;16(4):195-200
references.pmid: 9402009
references.citation: Development. 2012 Sep;139(17):3200-10
references.pmid: 22872087
references.citation: J Neurochem. 2000 Sep;75(3):991-1003
references.pmid: 10936180
references.citation: In Vitro Cell Dev Biol. 1987 Apr;23(4):239-46
references.pmid: 3553146
references.citation: J Neurosci. 2011 Aug 10;31(32):11678-91
references.pmid: 21832197
references.citation: J Biol Chem. 2009 Jul 24;284(30):20263-74
references.pmid: 19423701
references.citation: J Neurosci. 2003 May 15;23(10):4072-80
references.pmid: 12764094
references.citation: Brain Res Rev. 2007 Oct;55(2):204-19
references.pmid: 17467805
references.citation: J Neurosci. 1986 Dec;6(12):3721-30
references.pmid: 2947982
references.citation: Nat Cell Biol. 2000 Sep;2(9):574-81
references.pmid: 10980697
references.citation: Proc Natl Acad Sci U S A. 1986 May;83(10):3537-41
references.pmid: 3517863
references.citation: Cell Mol Life Sci. 2008 Nov;65(23):3809-21
references.pmid: 18791849
references.citation: Cell Adh Migr. 2011 Jan-Feb;5(1):52-5
references.pmid: 20953136
references.citation: Brain Res. 1984 Jun 8;302(2):323-34
references.pmid: 6329460
references.citation: Acta Biomater. 2012 Jan;8(1):165-72
references.pmid: 21911085
references.citation: J Neurocytol. 2003 Nov;32(9):1077-89
references.pmid: 15044840
references.citation: Endocr Rev. 1987 May;8(2):95-114
references.pmid: 2440668
references.citation: J Neurosci Res. 1998 Oct 15;54(2):233-47
references.pmid: 9788282
references.citation: Curr Opin Cell Biol. 2006 Oct;18(5):516-23
references.pmid: 16919435
references.citation: Eur J Neurosci. 1999 Nov;11(11):3777-88
references.pmid: 10583467
references.citation: Nat Rev Neurosci. 2007 Feb;8(2):141-51
references.pmid: 17237805
references.citation: J Biol Chem. 1997 Aug 22;272(34):21268-73
references.pmid: 9261137
references.citation: Biochem Biophys Res Commun. 2004 Jul 2;319(3):993-1000
references.pmid: 15184080
references.citation: J Alzheimers Dis. 2010;20(4):1069-82
references.pmid: 20413890
references.citation: Cell. 2003 Aug 22;114(4):469-82
references.pmid: 12941275
references.citation: J Cell Biol. 1989 Mar;108(3):1115-25
references.pmid: 2537833