title: In vitro models of axon regeneration.
abstract: A variety of in vitro models have been developed to understand the mechanisms underlying the regenerative failure of central nervous system (CNS) axons, and to guide pre-clinical development of regeneration-promoting therapeutics. These range from single-cell based assays that typically focus on molecular mechanisms to organotypic assays that aim to recapitulate in vivo behavior. By utilizing a combination of models, researchers can balance the speed, convenience, and mechanistic resolution of simpler models with the biological relevance of more complex models. This review will discuss a number of models that have been used to build our understanding of the molecular mechanisms of CNS axon regeneration.
authors.affiliation: Miami Project to Cure Paralysis, University of Miami Miller School of Medicine, Miami, FL 33136, USA.
authors.forename: Hassan
authors.initials: H
authors.lastname: Al-Ali
authors.affiliation: Miami Project to Cure Paralysis, University of Miami Miller School of Medicine, Miami, FL 33136, USA.
authors.forename: Samuel R
authors.initials: SR
authors.lastname: Beckerman
authors.affiliation: Miami Project to Cure Paralysis, University of Miami Miller School of Medicine, Miami, FL 33136, USA; Center for Computational Sciences, University of Miami Miller School of Medicine, Miami, FL 33136, USA; Neurological Surgery, University of Miami Miller School of Medicine, Miami, FL 33136, USA; Molecular & Cellular Pharmacology, University of Miami Miller School of Medicine, Miami, FL 33136, USA. Electronic address: jbixby@med.miami.edu.
authors.forename: John L
authors.initials: JL
authors.lastname: Bixby
authors.affiliation: Miami Project to Cure Paralysis, University of Miami Miller School of Medicine, Miami, FL 33136, USA; Center for Computational Sciences, University of Miami Miller School of Medicine, Miami, FL 33136, USA; Neurological Surgery, University of Miami Miller School of Medicine, Miami, FL 33136, USA. Electronic address: vlemmon@med.miami.edu.
authors.forename: Vance P
authors.initials: VP
authors.lastname: Lemmon
mesh_terms: D000818:Animals; D001369:Axons / Q000502:physiology*; D066298:In Vitro Techniques; D009416:Nerve Regeneration / Q000502:physiology*; D009474:Neurons / Q000166:cytology* / Q000502:physiology
keywords: Axon regeneration; Cell based assays; High content screening; In vitro models; Neurite outgrowth; Organotypic assays
references.citation: Sci Data. 2014 Nov 11;1:140038
references.pmid: 25977793
references.citation: J Neurosci Methods. 1997 Feb;71(2):143-55
references.pmid: 9128149
references.citation: Neuron. 2002 Jun 13;34(6):885-93
references.pmid: 12086637
references.citation: Development. 2003 Apr;130(8):1635-43
references.pmid: 12620987
references.citation: Neurosurg Clin N Am. 1991 Jan;2(1):31-42
references.pmid: 1821734
references.citation: Exp Neurol. 2015 Mar;265:152-9
references.pmid: 25585012
references.citation: Brain Res. 1979 Feb 23;162(2):273-9
references.pmid: 761090
references.citation: J Neurosci. 2003 May 1;23(9):3715-25
references.pmid: 12736342
references.citation: Proc Natl Acad Sci U S A. 1977 Oct;74(10):4516-9
references.pmid: 270699
references.citation: Nat Methods. 2005 Aug;2(8):599-605
references.pmid: 16094385
references.citation: Science. 2009 Oct 23;326(5952):592-6
references.pmid: 19833921
references.citation: PLoS One. 2015 Feb 25;10(2):e0118536
references.pmid: 25714396
references.citation: Neuron. 2006 Feb 2;49(3):365-77
references.pmid: 16446141
references.citation: Nat Neurosci. 2015 Jan;18(1):145-53
references.pmid: 25420068
references.citation: Cell. 2013 Nov 7;155(4):894-908
references.pmid: 24209626
references.citation: Mol Syst Biol. 2010 Jul;6:391
references.pmid: 20664637
references.citation: J Neurosci. 2011 Oct 5;31(40):14051-66
references.pmid: 21976490
references.citation: J Neurosci Methods. 2008 Jun 30;171(2):239-47
references.pmid: 18471889
references.citation: J Cell Physiol. 1965 Dec;66(3):325-33
references.pmid: 5884360
references.citation: Mol Cell Neurosci. 2010 May;44(1):43-54
references.pmid: 20159039
references.citation: PLoS One. 2012;7(5):e37574
references.pmid: 22629422
references.citation: Proc Natl Acad Sci U S A. 2011 Feb 15;108(7):3005-10
references.pmid: 21278334
references.citation: Proc Natl Acad Sci U S A. 1995 Aug 1;92(16):7287-91
references.pmid: 7638182
references.citation: Mol Cell Neurosci. 2003 Mar;22(3):319-30
references.pmid: 12691734
references.citation: Brain Res. 1982 May;256(1):59-66
references.pmid: 7093729
references.citation: J Neurosci Res. 1995 Dec;42(5):674-83
references.pmid: 8600300
references.citation: Exp Brain Res. 1988;73(2):393-401
references.pmid: 3265109
references.citation: J Neurosci. 2007 Dec 26;27(52):14494-501
references.pmid: 18160657
references.citation: J Neurocytol. 1984 Feb;13(1):165-82
references.pmid: 6707710
references.citation: J Neurosci. 1992 Apr;12(4):1253-61
references.pmid: 1372932
references.citation: Cell. 1995 Aug 11;82(3):359-70
references.pmid: 7634326
references.citation: Cell Transplant. 2013;22(4):571-617
references.pmid: 22944020
references.citation: J Neurosci. 1988 Feb;8(2):653-63
references.pmid: 3339432
references.citation: J Neurosci. 1984 Aug;4(8):1944-53
references.pmid: 6470762
references.citation: Development. 1987 Dec;101(4):909-13
references.pmid: 3503703
references.citation: Brain Res. 1976 Dec 17;118(2):299-303
references.pmid: 1000292
references.citation: J Cell Biol. 1977 Nov;75(2 Pt 1):464-74
references.pmid: 264120
references.citation: Assay Drug Dev Technol. 2015 Sep;13(7):377-88
references.pmid: 26230074
references.citation: Neuron. 1994 Sep;13(3):757-67
references.pmid: 7522484
references.citation: J Neurosci. 2003 Jul 2;23 (13):5393-406
references.pmid: 12843238
references.citation: Nat Neurosci. 2013 May;16(5):580-6
references.pmid: 23542688
references.citation: Neuron. 1999 May;23(1):83-91
references.pmid: 10402195
references.citation: Exp Brain Res. 1990;81(2):283-7
references.pmid: 2397757
references.citation: Biochem Biophys Res Commun. 2011 Feb 25;405(4):697-702
references.pmid: 21291862
references.citation: J Neurosci. 2012 May 2;32(18):6391-410
references.pmid: 22553043
references.citation: Proc Natl Acad Sci U S A. 2013 Mar 5;110(10):4045-50
references.pmid: 23431148
references.citation: J Neurosci. 1986 Feb;6(2):514-21
references.pmid: 2419526
references.citation: PLoS One. 2010 Jul 29;5(7):e11853
references.pmid: 20686615
references.citation: Brain Res. 1979 Jan 26;161(1):39-55
references.pmid: 758969
references.citation: J Neurobiol. 1992 Apr;23(3):322-36
references.pmid: 1624935
references.citation: Nature. 2014 Mar 13;507(7491):181-9
references.pmid: 24622198
references.citation: J Clin Invest. 1996 Jan 1;97(1):162-71
references.pmid: 8550829
references.citation: J Biomol Screen. 1999;4(2):67-73
references.pmid: 10838414
references.citation: Mol Cell Neurosci. 2001 Jul;18(1):26-43
references.pmid: 11461151
references.citation: Methods Enzymol. 2006;406:565-78
references.pmid: 16472688
references.citation: J Neurosci Methods. 2014 Oct 30;236:26-39
references.pmid: 25124852
references.citation: Surg Neurol. 2008 Jun;69(6):568-77; discussion 577
references.pmid: 18486695
references.citation: J Neurosci Res. 2004 Jul 15;77(2):299-307
references.pmid: 15211597
references.citation: Nat Protoc. 2010 Sep;5(10):1659-65
references.pmid: 20885378
references.citation: Neuron. 2010 Jun 10;66(5):663-70
references.pmid: 20547125
references.citation: Curr Chem Genom Transl Med. 2015 Feb 27;9:6-16
references.pmid: 25853055
references.citation: N Biotechnol. 2011 Jul;28(4):326-33
references.pmid: 21477670
references.citation: J Neurosci. 2007 Feb 14;27(7):1702-11
references.pmid: 17301178
references.citation: Mol Cell Neurosci. 2005 Aug;29(4):545-58
references.pmid: 15936953
references.citation: J Cell Biochem. 2004 Aug 1;92 (5):993-1003
references.pmid: 15258920
references.citation: Expert Opin Drug Discov. 2009 Oct 1;4(10):1035-1045
references.pmid: 20657751
references.citation: Nature. 2014 May 8;509(7499):189-94
references.pmid: 24776795
references.citation: Exp Neurol. 2013 Oct;248:170-82
references.pmid: 23792206
references.citation: Science. 1981 Nov 20;214(4523):931-3
references.pmid: 6171034
references.citation: Cancer Res. 1978 Nov;38(11 Pt 1):3751-7
references.pmid: 29704
references.citation: J Neurochem. 2007 Oct;103(1):181-9
references.pmid: 17608642
references.citation: Glia. 2012 May;60(6):894-907
references.pmid: 22374709
references.citation: J Neurosci. 2015 Jul 22;35(29):10429-39
references.pmid: 26203138
references.citation: Nature. 2000 Jan 27;403(6768):439-44
references.pmid: 10667797
references.citation: J Neurosci. 1987 Jan;7(1):201-12
references.pmid: 3543248
references.citation: J Neurosci Res. 1993 Aug 1;35(5):567-76
references.pmid: 8377226
references.citation: J Biol Chem. 2011 Aug 12;286(32):28011-8
references.pmid: 21685387
references.citation: Glia. 2014 Jan;62(1):78-95
references.pmid: 24272704
references.citation: Cold Spring Harb Protoc. 2013 Jun 01;2013(6):485-7
references.pmid: 23734014
references.citation: J Cell Biol. 1980 Jun;85(3):890-902
references.pmid: 6248568
references.citation: J Neurosci Res. 2006 Dec;84(8):1703-15
references.pmid: 17075918
references.citation: J Exp Zool. 1951 Mar;116(2):321-61
references.pmid: 14824426
references.citation: BioDrugs. 2010 Apr 1;24(2):99-108
references.pmid: 20199125
references.citation: Neuron. 2012 Jun 7;74(5):777-91
references.pmid: 22681683
references.citation: J Neurotrauma. 2015 Oct 1;32(19):1465-77
references.pmid: 25923828
references.citation: Methods Mol Biol. 2011;769:25-30
references.pmid: 21748666
references.citation: J Neurosci Res. 2008 Feb 15;86(3):581-92
references.pmid: 17918743
references.citation: PLoS One. 2011;6(6):e20857
references.pmid: 21698236
references.citation: Methods Mol Biol. 2012;814:327-40
references.pmid: 22144316
references.citation: Neuron. 1988 Nov;1(9):791-803
references.pmid: 2908449
references.citation: Exp Neurol. 2014 Mar;253:197-207
references.pmid: 24424280
references.citation: Glia. 2011 Jun;59(6):981-96
references.pmid: 21456043
references.citation: Nature. 2005 Jul 14;436(7048):193-200
references.pmid: 16015319
references.citation: Neuron. 2002 Feb 28;33(5):689-702
references.pmid: 11879647
references.citation: Brain. 2009 Nov;132(Pt 11):3102-21
references.pmid: 19783665
references.citation: J Neurosci. 1997 Aug 15;17(16):6256-63
references.pmid: 9236236
references.citation: Exp Neurol. 1990 Jul;109 (1):111-30
references.pmid: 2141574
references.citation: Mol Cell Neurosci. 2014 Mar;59:97-105
references.pmid: 24521823
references.citation: Langmuir. 2003 Mar 4;19(5):1551-1556
references.pmid: 20725530
references.citation: J Vis Exp. 2014 May 20;(87):null
references.pmid: 24894871
references.citation: J Neurosci. 2004 Jul 21;24(29):6531-9
references.pmid: 15269264
references.citation: Anal Biochem. 2006 Nov 1;358(1):1-10
references.pmid: 16979133
references.citation: Proc Natl Acad Sci U S A. 1954 Oct;40(10):1014-8
references.pmid: 16589582
references.citation: Science. 1982 May 21;216(4548):897-9
references.pmid: 7079743
references.citation: Neuroscience. 2005;134(2):449-65
references.pmid: 15964697
references.citation: Neuron. 1997 Mar;18(3):383-96
references.pmid: 9115733
references.citation: Proc Natl Acad Sci U S A. 2012 May 8;109(19):7517-22
references.pmid: 22529377
references.citation: Science. 2002 Feb 22;295(5559):1536-9
references.pmid: 11799202
references.citation: J Cell Biol. 2004 Jul 5;166(1):17-25
references.pmid: 15240567
references.citation: Nat Protoc. 2007;2(2):329-33
references.pmid: 17406593
references.citation: Science. 1979 Nov 30;206(4422):1079-80
references.pmid: 493992
references.citation: Invest Ophthalmol Vis Sci. 2002 Jun;43(6):2021-6
references.pmid: 12037014
references.citation: Cold Spring Harb Protoc. 2014 Aug 01;2014(8):826-38
references.pmid: 25086011
references.citation: Cell. 2010 Nov 12;143(4):527-39
references.pmid: 21074045
references.citation: J Neurosci. 1993 Apr;13(4):1784-92
references.pmid: 8385209
references.citation: Nat Protoc. 2007;2(5):1216-24
references.pmid: 17546017
references.citation: Nat Protoc. 2012 Sep;7(9):1741-54
references.pmid: 22936216
references.citation: Nat Rev Neurosci. 2003 Sep;4(9):703-13
references.pmid: 12951563
references.citation: Development. 1987 Dec;101(4):685-96
references.pmid: 3503693
references.citation: EMBO J. 1982;1(4):427-31
references.pmid: 6203734
references.citation: J Neurosci. 2003 Mar 1;23(5):1781-91
references.pmid: 12629182
references.citation: Proc Natl Acad Sci U S A. 1976 Jul;73(7):2424-8
references.pmid: 1065897
references.citation: Curr Chem Genomics. 2010 Dec 13;4:74-83
references.pmid: 21347208
references.citation: Development. 1995 May;121(5):1301-9
references.pmid: 7789262
references.citation: J Neurosci. 1991 Nov;11(11):3398-411
references.pmid: 1719160
references.citation: Nat Neurosci. 2004 Mar;7(3):261-8
references.pmid: 14770187
references.citation: J Neurosci Res. 2008 Aug 1;86(10):2214-26
references.pmid: 18438921
references.citation: J Neurosci. 1987 May;7(5):1595-600
references.pmid: 3572491
references.citation: Physiol Rev. 1996 Apr;76(2):319-70
references.pmid: 8618960
references.citation: Nat Protoc. 2012 Oct;7(10):1836-46
references.pmid: 22976355
references.citation: Invest Ophthalmol Vis Sci. 2010 Jan;51(1):526-34
references.pmid: 19628741
references.citation: Neuroinformatics. 2010 Jun;8(2):83-100
references.pmid: 20405243
references.citation: J Neurosci. 1988 Jul;8(7):2394-405
references.pmid: 3249232
references.citation: Neurosci Lett. 1978 Sep;9(2-3):97-104
references.pmid: 19605201
references.citation: J Neurochem. 2014 Jun;129(6):966-79
references.pmid: 24611815
references.citation: Cell Death Dis. 2013 Jul 18;4:e734
references.pmid: 23868067
references.citation: Cell. 1999 Oct 29;99(3):323-34
references.pmid: 10555148
references.citation: J Neurosci. 1988 Apr;8(4):1454-68
references.pmid: 3282038
references.citation: Proc Natl Acad Sci U S A. 1976 Feb;73(2):410-4
references.pmid: 1061144
references.citation: Dev Biol. 1989 Oct;135(2):449-58
references.pmid: 2776978
references.citation: Nature. 2011 May 12;473(7346):221-5
references.pmid: 21490598
references.citation: Curr Biol. 2005 Sep 20;15(18):R749-53
references.pmid: 16169471
references.citation: Brain Res. 1977 May 13;126(3):397-42
references.pmid: 861729
references.citation: J Cell Biol. 2002 May 13;157(4):565-70
references.pmid: 12011108
references.citation: Mol Cell Neurosci. 2001 Jun;17(6):1048-58
references.pmid: 11414793
references.citation: Science. 1998 Sep 4;281(5382):1515-8
references.pmid: 9727979
references.citation: Assay Drug Dev Technol. 2014 Nov-Dec;12(9-10):536-47
references.pmid: 25506803
references.citation: J Neurosci. 2001 Jul 1;21(13):4731-9
references.pmid: 11425900
references.citation: J Neurotrauma. 1995 Jun;12(3):325-39
references.pmid: 7473807
references.citation: J Vis Exp. 2009 May 05;(27):null
references.pmid: 19417729
references.citation: Science. 1976 Apr 16;192(4236):218-26
references.pmid: 769162
references.citation: Science. 2012 Jul 20;337(6092):358-62
references.pmid: 22745251
references.citation: Nat Protoc. 2006;1(3):1322-8
references.pmid: 17406418
references.citation: Brain Res Brain Res Protoc. 2002 Oct;10(2):60-8
references.pmid: 12431704
references.citation: Mol Cell Neurosci. 2000 Nov;16(5):686-95
references.pmid: 11083928
references.citation: Nat Rev Neurosci. 2006 Aug;7(8):617-27
references.pmid: 16858390
references.citation: ACS Chem Biol. 2015 Aug 21;10(8):1939-51
references.pmid: 26056718
references.citation: J Neurosci. 1999 Dec 15;19(24):10778-88
references.pmid: 10594061
references.citation: J Neurosci. 2011 Aug 31;31(35):12471-82
references.pmid: 21880909
references.citation: Neuron. 1995 Oct;15(4):805-19
references.pmid: 7576630
references.citation: ACS Chem Biol. 2013 May 17;8(5):1027-36
references.pmid: 23480631
references.citation: Science. 2008 Nov 7;322(5903):963-6
references.pmid: 18988856
references.citation: J Neurosci. 2010 Mar 31;30(13):4693-706
references.pmid: 20357120
references.citation: Front Mol Neurosci. 2012 Mar 01;5:24
references.pmid: 22403528
references.citation: J Neurosci. 1992 Mar;12(3):818-26
references.pmid: 1545241
references.citation: Glia. 1991;4(1):46-55
references.pmid: 1828786
references.citation: Nat Rev Neurosci. 2009 Mar;10(3):235-41
references.pmid: 19229242
references.citation: Cell. 1994 Aug 12;78(3):409-24
references.pmid: 8062384
references.citation: Neuron. 1999 Jun;23(2):297-308
references.pmid: 10399936
references.citation: Nature. 1987 Nov 19-25;330(6145):254-6
references.pmid: 3313064
references.citation: Neurotoxicol Teratol. 2004 May-Jun;26(3):397-406
references.pmid: 15113601
references.citation: Cell Death Differ. 2008 Feb;15(2):283-98
references.pmid: 18007665
references.citation: J Neurogenet. 2011 Oct;25(3):88-103
references.pmid: 21797804
references.citation: Development. 1991 Dec;113(4):1473-85
references.pmid: 1811954
references.citation: Annu Rev Neurosci. 2007;30:209-33
references.pmid: 17341159
references.citation: Science. 2005 Oct 7;310(5745):106-10
references.pmid: 16210539
references.citation: J Neurobiol. 1994 Jun;25(6):599-611
references.pmid: 8071664
references.citation: Nature. 2000 Jan 6;403(6765):93-8
references.pmid: 10638760
references.citation: Nat Protoc. 2014 May;9(5):1028-37
references.pmid: 24705599
references.citation: Spine (Phila Pa 1976). 2009 Nov 1;34(23 ):2494-9
references.pmid: 19927097
references.citation: Neuron. 2003 May 8;38(3):403-16
references.pmid: 12741988
references.citation: Nature. 2001 Jan 18;409(6818):341-6
references.pmid: 11201742
references.citation: Dev Biol. 1983 Jan;95(1):56-64
references.pmid: 6825932
references.citation: Philos Trans R Soc Lond B Biol Sci. 2006 Sep 29;361(1473):1565-74
references.pmid: 16939975
references.citation: Science. 2009 Oct 9;326(5950):298-301
references.pmid: 19815778
references.citation: Nat Rev Neurol. 2013 Jan;9(1):44-53
references.pmid: 23165340
references.citation: Exp Neurol. 1991 Jun;112(3):243-51
references.pmid: 2029925
references.citation: Nat Protoc. 2007;2(1):152-60
references.pmid: 17401349
references.citation: Tissue Eng Part A. 2010 Oct;16(10):3173-84
references.pmid: 20649441
references.citation: Neuron. 1997 Sep;19(3):575-89
references.pmid: 9331350
references.citation: Neuroscience. 2005;132(1):87-102
references.pmid: 15780469
references.citation: J Exp Biol. 1981 Dec;95:231-40
references.pmid: 7334319
references.citation: Science. 1996 May 24;272(5265):1182-7
references.pmid: 8638165
references.citation: Glia. 2008 Nov 15;56(15):1691-709
references.pmid: 18618667
references.citation: Exp Neurol. 2010 May;223(1):45-50
references.pmid: 20079353
references.citation: Hum Mol Genet. 2010 Apr 15;19(R1):R71-6
references.pmid: 20418487
references.citation: J Neurosci. 2010 Aug 11;30(32):10899-904
references.pmid: 20702718