title: Controlled induction of human pancreatic progenitors produces functional beta-like cells in vitro.
abstract: Directed differentiation of human pluripotent stem cells into functional insulin-producing beta-like cells holds great promise for cell replacement therapy for patients suffering from diabetes. This approach also offers the unique opportunity to study otherwise inaccessible aspects of human beta cell development and function in vitro. Here, we show that current pancreatic progenitor differentiation protocols promote precocious endocrine commitment, ultimately resulting in the generation of non-functional polyhormonal cells. Omission of commonly used BMP inhibitors during pancreatic specification prevents precocious endocrine formation while treatment with retinoic acid followed by combined EGF/KGF efficiently generates both PDX1(+) and subsequent PDX1(+)/NKX6.1(+) pancreatic progenitor populations, respectively. Precise temporal activation of endocrine differentiation in PDX1(+)/NKX6.1(+) progenitors produces glucose-responsive beta-like cells in vitro that exhibit key features of bona fide human beta cells, remain functional after short-term transplantation, and reduce blood glucose levels in diabetic mice. Thus, our simplified and scalable system accurately recapitulates key steps of human pancreas development and provides a fast and reproducible supply of functional human beta-like cells.
authors.affiliation: Diabetes Center, University of California San Francisco, San Francisco, CA, USA.
authors.forename: Holger A
authors.initials: HA
authors.lastname: Russ
authors.affiliation: Diabetes Center, University of California San Francisco, San Francisco, CA, USA.
authors.forename: Audrey V
authors.initials: AV
authors.lastname: Parent
authors.affiliation: Diabetes Center, University of California San Francisco, San Francisco, CA, USA.
authors.forename: Jennifer J
authors.initials: JJ
authors.lastname: Ringler
authors.affiliation: Diabetes Center, University of California San Francisco, San Francisco, CA, USA.
authors.forename: Thomas G
authors.initials: TG
authors.lastname: Hennings
authors.affiliation: Diabetes Center, University of California San Francisco, San Francisco, CA, USA.
authors.forename: Gopika G
authors.initials: GG
authors.lastname: Nair
authors.affiliation: Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Center for Reproductive Sciences and Department of Urology, University of California San Francisco, San Francisco, CA, USA.
authors.forename: Mayya
authors.initials: M
authors.lastname: Shveygert
authors.affiliation: Diabetes Center, University of California San Francisco, San Francisco, CA, USA.
authors.forename: Tingxia
authors.initials: T
authors.lastname: Guo
authors.affiliation: Diabetes Center, University of California San Francisco, San Francisco, CA, USA.
authors.forename: Sapna
authors.initials: S
authors.lastname: Puri
authors.affiliation: Division of Metabolism, Endocrinology & Diabetes, University of Michigan Medical School, Brehm Tower Ann Arbor, MI, USA.
authors.forename: Leena
authors.initials: L
authors.lastname: Haataja
authors.affiliation: Diabetes and Obesity Center of Excellence, Department of Medicine, Institute for Stem Cells and Regenerative Medicine, University of Washington, Seattle, WA, USA.
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Cirulli
authors.affiliation: Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Center for Reproductive Sciences and Department of Urology, University of California San Francisco, San Francisco, CA, USA.
authors.forename: Robert
authors.initials: R
authors.lastname: Blelloch
authors.affiliation: Diabetes Center, University of California San Francisco, San Francisco, CA, USA.
authors.forename: Greg L
authors.initials: GL
authors.lastname: Szot
authors.affiliation: Division of Metabolism, Endocrinology & Diabetes, University of Michigan Medical School, Brehm Tower Ann Arbor, MI, USA.
authors.forename: Peter
authors.initials: P
authors.lastname: Arvan
authors.affiliation: Diabetes Center, University of California San Francisco, San Francisco, CA, USA mhebrok@diabetes.ucsf.edu.
authors.forename: Matthias
authors.initials: M
authors.lastname: Hebrok
mesh_terms: D000818:Animals; D001786:Blood Glucose / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002478:Cells, Cultured; D003921:Diabetes Mellitus, Experimental / Q000097:blood / Q000628:therapy; D053595:Embryonic Stem Cells / Q000166:cytology / Q000502:physiology*; D005947:Glucose / Q000494:pharmacology; D006801:Humans; D050417:Insulin-Secreting Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology* / Q000637:transplantation; D051379:Mice; D016513:Mice, SCID; D008822:Mice, Transgenic; D010179:Pancreas / Q000166:cytology*; D013311:Streptozocin
chemical_list: D001786:Blood Glucose; D013311:Streptozocin; D005947:Glucose
keywords: beta‐like cells; diabetes; human embryonic stem cells; insulin‐producing cells, pancreas