title: Efficient generation of lens progenitor cells and lentoid bodies from human embryonic stem cells in chemically defined conditions.
abstract: The eye lens is an encapsulated avascular organ whose function is to focus light on the retina. Lens comprises a single progenitor cell lineage in multiple states of differentiation. Disruption of lens function leading to protein aggregation and opacity results in age-onset cataract. Cataract is a complex disease involving genetic and environmental factors. Here, we report the development of a new 3-stage system that differentiates human embryonic stem cells (hESCs) into large quantities of lens progenitor-like cells and differentiated 3-dimensional lentoid bodies. Inhibition of BMP signaling by noggin triggered differentiation of hESCs toward neuroectoderm. Subsequent reactivation of BMP and activation of FGF signaling stimulated formation of lens progenitor cells marked by the expression of PAX6 and alpha-crystallins. The formation of lentoid bodies was most efficient in the presence of FGF2 and Wnt-3a, yielding approximately 1000 lentoid bodies/30-mm well. Lentoid bodies expressed and accumulated lens-specific markers including alphaA-, alphaB-, beta-, and gamma-crystallins, filensin, CP49, and MIP/aquaporin 0. Collectively, these studies identify a novel procedure to generate lens cells from hESCs that can be applied for studies of lens differentiation and cataractogenesis using induced pluripotent stem (iPS) cells derived from various cataract patients.
authors.affiliation: Department of Ophthalmology, Albert Einstein College of Medicine, 1300 Morris Park Ave., Bronx, NY 10461, USA.
authors.forename: Chunbo
authors.initials: C
authors.lastname: Yang
authors.forename: Ying
authors.initials: Y
authors.lastname: Yang
authors.forename: Lisa
authors.initials: L
authors.lastname: Brennan
authors.forename: Eric E
authors.initials: EE
authors.lastname: Bouhassira
authors.forename: Marc
authors.initials: M
authors.lastname: Kantorow
authors.forename: Ales
authors.initials: A
authors.lastname: Cvekl
mesh_terms: D000328:Adult; D015153:Blotting, Western; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology; D007908:Lens, Crystalline / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D008297:Male; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission; D008875:Middle Aged; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure
chemical_list: D036341:Intercellular Signaling Peptides and Proteins
references.citation: Exp Eye Res. 2009 Feb;88(2):133-9
references.pmid: 18840431
references.citation: Development. 2002 Aug;129(15):3727-37
references.pmid: 12117821
references.citation: Cell Stem Cell. 2009 Oct 2;5(4):420-33
references.pmid: 19796622
references.citation: Exp Eye Res. 2009 Feb;88(2):195-203
references.pmid: 18588875
references.citation: Genes Dev. 1995 Nov 15;9(22):2808-20
references.pmid: 7590255
references.citation: Development. 2004 Apr;131(8):1813-24
references.pmid: 15084465
references.citation: Cell Stem Cell. 2009 Aug 7;5(2):126-8
references.pmid: 19664982
references.citation: Exp Eye Res. 1991 Jun;52(6):647-59
references.pmid: 1855541
references.citation: Dev Biol. 2005 Apr 1;280(1):1-14
references.pmid: 15766743
references.citation: Proc Natl Acad Sci U S A. 2006 Aug 22;103(34):12769-74
references.pmid: 16908856
references.citation: Dev Biol. 2008 Dec 15;324(2):202-12
references.pmid: 18848538
references.citation: Proc Natl Acad Sci U S A. 2006 May 2;103(18):6907-12
references.pmid: 16632596
references.citation: Nat Rev Neurosci. 2002 Apr;3(4):271-80
references.pmid: 11967557
references.citation: Science. 2007 Dec 21;318(5858):1917-20
references.pmid: 18029452
references.citation: Semin Cell Dev Biol. 2006 Dec;17(6):726-40
references.pmid: 17116415
references.citation: Invest Ophthalmol Vis Sci. 2007 Mar;48(3):1245-52
references.pmid: 17325169
references.citation: Development. 1989 Oct;107(2):221-8
references.pmid: 2632221
references.citation: Development. 2001 Oct;128(20):3995-4010
references.pmid: 11641223
references.citation: Arch Ophthalmol. 2007 Feb;125(2):165-73
references.pmid: 17296892
references.citation: Cell. 2006 Aug 25;126(4):663-76
references.pmid: 16904174
references.citation: Cell Stem Cell. 2009 Oct 2;5(4):396-408
references.pmid: 19796620
references.citation: Genes Dev. 1998 Dec 1;12(23):3764-75
references.pmid: 9851982
references.citation: Nature. 1995 Jul 27;376(6538):331-3
references.pmid: 7630398
references.citation: Invest Ophthalmol Vis Sci. 2003 Jun;44(6):2689-93
references.pmid: 12766074
references.citation: Mech Dev. 2000 May;93(1-2):195-200
references.pmid: 10781956
references.citation: Dev Biol. 2004 Dec 1;276(1):1-15
references.pmid: 15531360
references.citation: Stem Cells. 2009 Jun;27(6):1298-308
references.pmid: 19489095
references.citation: Int J Dev Biol. 2004;48(8-9):783-91
references.pmid: 15558471
references.citation: Dev Biol. 2008 Jun 15;318(2):276-88
references.pmid: 18455718
references.citation: Dev Dyn. 1996 Sep;207(1):75-88
references.pmid: 8875078
references.citation: Nature. 2009 Nov 19;462(7271):315-22
references.pmid: 19829295
references.citation: Dev Biol. 2010 Jan 15;337(2):195-8
references.pmid: 19913010
references.citation: Invest Ophthalmol Vis Sci. 2007 Dec;48(12):5638-46
references.pmid: 18055814
references.citation: EMBO J. 2006 Nov 15;25(22):5383-95
references.pmid: 17066077
references.citation: Mol Vis. 2001 Jun 25;7:138-44
references.pmid: 11436000
references.citation: Development. 2005 Sep;132(18):4051-62
references.pmid: 16093325
references.citation: Prog Retin Eye Res. 2007 Nov;26(6):555-97
references.pmid: 17905638
references.citation: Dev Biol. 2003 Jul 1;259(1):48-61
references.pmid: 12812787
references.citation: J Neurosci. 1995 Sep;15(9):6077-84
references.pmid: 7666191
references.citation: Stem Cells. 2007 Oct;25(10):2567-74
references.pmid: 17641388
references.citation: Genes Dev. 2005 May 15;19(10):1129-55
references.pmid: 15905405
references.citation: Invest Ophthalmol Vis Sci. 1996 Oct;37(11):2276-84
references.pmid: 8843924
references.citation: Nature. 2003 Aug 28;424(6952):1071-4
references.pmid: 12944971
references.citation: Dev Biol. 2009 Nov 15;335(2):305-16
references.pmid: 19733164
references.citation: BMC Dev Biol. 2008 Sep 19;8:88
references.pmid: 18803847
references.citation: Dev Biol. 2008 Dec 1;324(1):161-76
references.pmid: 18824165
references.citation: Development. 1993 May;118(1):117-26
references.pmid: 7690700
references.citation: J Cell Sci. 1995 Apr;108 ( Pt 4):1397-406
references.pmid: 7615661
references.citation: Nat Biotechnol. 2008 Feb;26(2):215-24
references.pmid: 18246062
references.citation: Invest Ophthalmol. 1974 Dec;13(12):973-81
references.pmid: 4430579