title: Regulation of prenatal human retinal neurosphere growth and cell fate potential by retinal pigment epithelium and Mash1.
abstract: During development of the central nervous system, stem and progenitor cell proliferation and differentiation are controlled by complex inter- and intracellular interactions that orchestrate the precise spatiotemporal production of particular cell types. Within the embryonic retina, progenitor cells are located adjacent to the retinal pigment epithelium (RPE), which differentiates prior to the neurosensory retina and has the capacity to secrete a multitude of growth factors. We found that secreted proteinaceous factors in human prenatal RPE conditioned medium (RPE CM) prolonged and enhanced the growth of human prenatal retinal neurospheres. The growth-promoting activity of RPE CM was mitogen-dependent and associated with an acute increase in transcription factor phosphorylation. Expanded populations of RPE CM-treated retinal neurospheres expressed numerous neurodevelopmental and eye specification genes and markers characteristic of neural and retinal progenitor cells, but gradually lost the potential to generate neurons upon differentiation. Misexpression of Mash1 restored the neurogenic potential of long-term cultures, yielding neurons with phenotypic characteristics of multiple inner retinal cell types. Thus, a novel combination of extrinsic and intrinsic factors was required to promote both progenitor cell proliferation and neuronal multipotency in human retinal neurosphere cultures. These results support a pro-proliferative and antiapoptotic role for RPE in human retinal development, reveal potential limitations of human retinal progenitor culture systems, and suggest a means for overcoming cell fate restriction in vitro.
authors.affiliation: Department of Ophthalmology and Visual Sciences, University of Wisconsin, Madison, Wisconsin 53705, USA. dgamm@wisc.edu
authors.forename: David M
authors.initials: DM
authors.lastname: Gamm
authors.forename: Lynda S
authors.initials: LS
authors.lastname: Wright
authors.forename: Elizabeth E
authors.initials: EE
authors.lastname: Capowski
authors.forename: Rebecca L
authors.initials: RL
authors.lastname: Shearer
authors.forename: Jason S
authors.initials: JS
authors.lastname: Meyer
authors.forename: Hyun-Jung
authors.initials: HJ
authors.lastname: Kim
authors.forename: Bernard L
authors.initials: BL
authors.lastname: Schneider
authors.forename: John Nicholas
authors.initials: JN
authors.lastname: Melvan
authors.forename: Clive N
authors.initials: CN
authors.lastname: Svendsen
mesh_terms: D017209:Apoptosis; D051792:Basic Helix-Loop-Helix Proteins / Q000378:metabolism*; D002454:Cell Differentiation; D019070:Cell Lineage; D049109:Cell Proliferation; D017077:Culture Media, Conditioned / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D009474:Neurons / Q000378:metabolism*; D010641:Phenotype; D010766:Phosphorylation; D012160:Retina / Q000166:cytology; D055213:Retinal Pigment Epithelium / Q000378:metabolism*; D013234:Stem Cells / Q000166:cytology; D014157:Transcription Factors / Q000378:metabolism
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D017077:Culture Media, Conditioned; D014157:Transcription Factors; C065500:ASCL1 protein, human
references.citation: J Neurosci Methods. 1998 Dec 1;85(2):141-52
references.pmid: 9874150
references.citation: Curr Opin Neurobiol. 1999 Feb;9(1):37-46
references.pmid: 10072376
references.citation: J Neurobiol. 2004 Dec;61(3):359-76
references.pmid: 15452852
references.citation: Int J Dev Biol. 2004;48(8-9):1003-14
references.pmid: 15558491
references.citation: Br J Ophthalmol. 2005 Jan;89(1):102-6
references.pmid: 15615756
references.citation: Development. 2005 Jan;132(2):311-22
references.pmid: 15604100
references.citation: Dev Biol. 2005 Mar 15;279(2):308-21
references.pmid: 15733660
references.citation: Stem Cells. 2005 Mar;23(3):424-32
references.pmid: 15749937
references.citation: Exp Eye Res. 2005 Apr;80(4):515-25
references.pmid: 15781279
references.citation: Semin Ophthalmol. 2005 Jan-Mar;20(1):25-30
references.pmid: 15804841
references.citation: Semin Cell Dev Biol. 2005 Jun;16(3):407-21
references.pmid: 15840449
references.citation: Dev Neurosci. 2004;26(5-6):294-307
references.pmid: 15855758
references.citation: Exp Cell Res. 2005 Jun 10;306(2):343-8
references.pmid: 15925590
references.citation: Neuron. 2005 Jun 2;46(5):731-44
references.pmid: 15924860
references.citation: Ann N Y Acad Sci. 2005 May;1049:107-17
references.pmid: 15965111
references.citation: Dev Biol. 2005 Aug 15;284(2):464-78
references.pmid: 16038893
references.citation: Nat Neurosci. 2006 Mar;9(3):331-9
references.pmid: 16491078
references.citation: Genes Dev. 2006 May 1;20(9):1187-202
references.pmid: 16651659
references.citation: Exp Cell Res. 2006 Jul 1;312(11):2107-20
references.pmid: 16631163
references.citation: Biochemistry (Mosc). 2006 Jul;71(7):767-74
references.pmid: 16903831
references.citation: Brain Res Rev. 2006 Sep;52(2):264-74
references.pmid: 16764935
references.citation: Stem Cells. 2006 Sep;24(9):2060-70
references.pmid: 16644923
references.citation: Pigment Cell Res. 2006 Oct;19(5):380-94
references.pmid: 16965267
references.citation: Trends Neurosci. 2006 Oct;29(10):563-70
references.pmid: 16920202
references.citation: Dev Biol. 2006 Nov 1;299(1):283-302
references.pmid: 16949068
references.citation: Methods Enzymol. 2006;419:52-73
references.pmid: 17141051
references.citation: Proc Natl Acad Sci U S A. 2006 Dec 12;103(50):18998-9003
references.pmid: 17148603
references.citation: Mol Vis. 2006;12:1606-14
references.pmid: 17200660
references.citation: Exp Neurol. 2007 Feb;203(2):394-405
references.pmid: 17034791
references.citation: Stem Cells. 2007 Aug;25(8):2033-43
references.pmid: 17525239
references.citation: Prog Retin Eye Res. 2003 Sep;22(5):567-77
references.pmid: 12892642
references.citation: Brain Res Dev Brain Res. 2003 Oct 10;145(1):1-8
references.pmid: 14519488
references.citation: DNA Cell Biol. 2003 Oct;22(10):607-20
references.pmid: 14611682
references.citation: J Neurosci. 2007 Aug 8;27(32):8654-64
references.pmid: 17687043
references.citation: Dev Biol. 1997 Aug 15;188(2):267-79
references.pmid: 9268574
references.citation: Eur J Neurosci. 1997 Sep;9(9):1795-803
references.pmid: 9383202
references.citation: Brain Res Dev Brain Res. 1998 Apr 17;107(1):57-69
references.pmid: 9602059
references.citation: J Neurosci. 1998 Oct 1;18(19):7869-80
references.pmid: 9742155
references.citation: Exp Eye Res. 2007 Nov;85(5):587-601
references.pmid: 17845803
references.citation: Brain Res. 2007 Dec 14;1185:59-67
references.pmid: 17961525
references.citation: Hum Gene Ther. 2000 Jan 1;11(1):179-90
references.pmid: 10646649
references.citation: J Comp Neurol. 2000 May 15;420(4):437-44
references.pmid: 10805919
references.citation: Neuron. 2000 May;26(2):383-94
references.pmid: 10839357
references.citation: Development. 2000 Jul;127(14):3021-30
references.pmid: 10862740
references.citation: Development. 2000 Aug;127(15):3237-47
references.pmid: 10887080
references.citation: J Neurosci. 2000 Oct 1;20(19):7149-57
references.pmid: 11007870
references.citation: Development. 2001 Jan;128(1):95-105
references.pmid: 11092815
references.citation: Genesis. 2000 Nov-Dec;28(3-4):135-42
references.pmid: 11105055
references.citation: Proc Natl Acad Sci U S A. 2000 Dec 19;97(26):14720-5
references.pmid: 11121071
references.citation: Nat Neurosci. 2001 Mar;4(3):247-52
references.pmid: 11224540
references.citation: Nat Rev Neurosci. 2001 Feb;2(2):109-18
references.pmid: 11252990
references.citation: Nat Biotechnol. 2001 May;19(5):475-9
references.pmid: 11329020
references.citation: Brain Res Dev Brain Res. 2001 Apr 30;127(2):185-7
references.pmid: 11335005
references.citation: Neuroreport. 2001 May 25;12(7):A39-46
references.pmid: 11388446
references.citation: Prog Brain Res. 2001;131:541-54
references.pmid: 11420969
references.citation: Nature. 2001 Nov 1;414(6859):112-7
references.pmid: 11689956
references.citation: J Anat. 2002 Jan;200(Pt 1):51-6
references.pmid: 11833654
references.citation: Development. 2002 May;129(9):2283-91
references.pmid: 11959835
references.citation: Development. 2002 Oct;129(19):4435-42
references.pmid: 12223402
references.citation: Exp Neurol. 2002 Sep;177(1):326-31
references.pmid: 12429235
references.citation: Development. 2003 Jan;130(1):185-95
references.pmid: 12441302
references.citation: Nat Neurosci. 2002 Dec;5(12):1271-8
references.pmid: 12426573
references.citation: J Comp Neurol. 2003 Jun 23;461(2):187-204
references.pmid: 12724837
references.citation: Eur J Neurosci. 2003 Apr;17(8):1542-8
references.pmid: 12752371
references.citation: Glia. 2003 Jul;43(1):70-6
references.pmid: 12761869
references.citation: Clin Experiment Ophthalmol. 2003 Jun;31(3):246-9
references.pmid: 12786777
references.citation: Lab Invest. 2003 Jul;83(7):949-62
references.pmid: 12861035
references.citation: Nat Rev Neurosci. 2003 Aug;4(8):628-36
references.pmid: 12894238
references.citation: In Vitro Cell Dev Biol Anim. 2007 Nov-Dec;43(10):361-70
references.pmid: 17924175
references.citation: Invest Ophthalmol Vis Sci. 2008 Feb;49(2):788-99
references.pmid: 18235029
references.citation: Curr Eye Res. 2008 Feb;33(2):185-91
references.pmid: 18293190
references.citation: Neuron. 2003 Dec 4;40(5):897-904
references.pmid: 14659089
references.citation: Semin Cell Dev Biol. 2004 Feb;15(1):83-9
references.pmid: 15036211
references.citation: Semin Cell Dev Biol. 2004 Feb;15(1):91-103
references.pmid: 15036212
references.citation: Exp Eye Res. 2004 May;78(5):1025-36
references.pmid: 15051483
references.citation: Vis Neurosci. 2003 Nov-Dec;20(6):589-600
references.pmid: 15088712
references.citation: J Comp Neurol. 2004 Jun 21;474(2):304-24
references.pmid: 15164429
references.citation: J Neurosci Res. 2004 Aug 1;77(3):334-43
references.pmid: 15248289
references.citation: Curr Eye Res. 2004 May;28(5):327-36
references.pmid: 15287369
references.citation: Prog Retin Eye Res. 2004 Sep;23(5):561-77
references.pmid: 15302351
references.citation: Invest Ophthalmol Vis Sci. 2004 Nov;45(11):4167-73
references.pmid: 15505071
references.citation: Am J Ophthalmol. 1970 Jun;69(6):1032-46
references.pmid: 5423768
references.citation: J Neurosci. 1986 Jul;6(7):1885-96
references.pmid: 3525774
references.citation: Anat Rec. 1985 Jun;212(2):199-205
references.pmid: 3842042
references.citation: Nature. 1987 Jul 9-15;328(6126):131-6
references.pmid: 3600789
references.citation: Science. 1992 Mar 27;255(5052):1707-10
references.pmid: 1553558
references.citation: Exp Eye Res. 1992 Feb;54(2):193-200
references.pmid: 1559548
references.citation: Exp Eye Res. 1992 May;54(5):645-59
references.pmid: 1385580
references.citation: Brain Res Dev Brain Res. 1994 Jul 15;80(1-2):81-95
references.pmid: 7955364
references.citation: Invest Ophthalmol Vis Sci. 1995 Jun;36(7):1280-9
references.pmid: 7775105
references.citation: Genes Dev. 1995 Dec 15;9(24):3136-48
references.pmid: 8543157
references.citation: Proc Natl Acad Sci U S A. 1996 Jan 23;93(2):589-95
references.pmid: 8570600
references.citation: Curr Biol. 1995 Nov 1;5(11):1286-95
references.pmid: 8574586
references.citation: J Neurosci Res. 1996 Jun 15;44(6):519-31
references.pmid: 8794943
references.citation: Brain Res Dev Brain Res. 1996 May 31;93(1-2):88-99
references.pmid: 8804695
references.citation: Genes Cells. 1996 Aug;1(8):765-74
references.pmid: 9077445
references.citation: Proc Natl Acad Sci U S A. 1997 Apr 1;94(7):3088-93
references.pmid: 9096350
references.citation: Trends Neurosci. 1997 Apr;20(4):165-9
references.pmid: 9106357