title: Stage-specific signaling through TGFβ family members and WNT regulates patterning and pancreatic specification of human pluripotent stem cells.
abstract: The generation of insulin-producing β-cells from human pluripotent stem cells is dependent on efficient endoderm induction and appropriate patterning and specification of this germ layer to a pancreatic fate. In this study, we elucidated the temporal requirements for TGFβ family members and canonical WNT signaling at these developmental stages and show that the duration of nodal/activin A signaling plays a pivotal role in establishing an appropriate definitive endoderm population for specification to the pancreatic lineage. WNT signaling was found to induce a posterior endoderm fate and at optimal concentrations enhanced the development of pancreatic lineage cells. Inhibition of the BMP signaling pathway at specific stages was essential for the generation of insulin-expressing cells and the extent of BMP inhibition required varied widely among the cell lines tested. Optimal stage-specific manipulation of these pathways resulted in a striking 250-fold increase in the levels of insulin expression and yielded populations containing up to 25% C-peptide+ cells.
authors.affiliation: McEwen Centre for Regenerative Medicine, University Health Network, Toronto, Ontario, Canada.
authors.forename: M Cristina
authors.initials: MC
authors.lastname: Nostro
authors.forename: Farida
authors.initials: F
authors.lastname: Sarangi
authors.forename: Shinichiro
authors.initials: S
authors.lastname: Ogawa
authors.forename: Audrey
authors.initials: A
authors.lastname: Holtzinger
authors.forename: Barbara
authors.initials: B
authors.lastname: Corneo
authors.forename: Xueling
authors.initials: X
authors.lastname: Li
authors.forename: Suzanne J
authors.initials: SJ
authors.lastname: Micallef
authors.forename: In-Hyun
authors.initials: IH
authors.lastname: Park
authors.forename: Christina
authors.initials: C
authors.lastname: Basford
authors.forename: Michael B
authors.initials: MB
authors.lastname: Wheeler
authors.forename: George Q
authors.initials: GQ
authors.lastname: Daley
authors.forename: Andrew G
authors.initials: AG
authors.lastname: Elefanty
authors.forename: Edouard G
authors.initials: EG
authors.lastname: Stanley
authors.forename: Gordon
authors.initials: G
authors.lastname: Keller
mesh_terms: D028341:Activins / Q000378:metabolism; D019521:Body Patterning; D019485:Bone Morphogenetic Proteins / Q000037:antagonists & inhibitors; D002096:C-Peptide; D002460:Cell Line; D019070:Cell Lineage; D004707:Endoderm; D006801:Humans; D007328:Insulin / Q000096:biosynthesis; D050417:Insulin-Secreting Cells / Q000166:cytology*; D010179:Pancreas / Q000166:cytology*; D039904:Pluripotent Stem Cells / Q000502:physiology*; D015398:Signal Transduction / Q000502:physiology; D016212:Transforming Growth Factor beta / Q000378:metabolism*; D051153:Wnt Proteins / Q000378:metabolism*
chemical_list: D019485:Bone Morphogenetic Proteins; D002096:C-Peptide; D007328:Insulin; D016212:Transforming Growth Factor beta; D051153:Wnt Proteins; C422405:activin A; D028341:Activins