title: Directed differentiation of human embryonic stem cells into the pancreatic endocrine lineage.
abstract: Human embryonic stem (hES) cells represent a potentially unlimited source of transplantable beta-cells for the treatment of diabetes. Here we describe a differentiation strategy that reproducibly directs HES3, an National Institutes of Health (NIH)-registered hES cell line, into cells of the pancreatic endocrine lineage. HES3 cells are removed from their feeder layer and cultured as embryoid bodies in a three-dimensional matrix in the presence of Activin A and Bmp4 to induce definitive endoderm. Next, growth factors known to promote the proliferation and differentiation of pancreatic ductal epithelial cells to glucose-sensing, insulin-secreting beta-cells are added. Pdx1 expression, which identifies pancreatic progenitors, is detected as early as day 12 of differentiation. By day 34, Pdx1+ cells comprise between 5% and 20% of the total cell population and Insulin gene expression is up-regulated, with release of C-peptide into the culture medium. Unlike another recent report of the induction of insulin+ cells in differentiated hES cell populations, we are unable to detect the expression of other pancreatic hormones in insulin+ cells. When transplanted into severe combined immunodeficiency (SCID) mice, differentiated cell populations retain their endocrine identity and synthesize insulin.
authors.affiliation: ES Cell International Pte, Ltd., Singapore 138667.
authors.forename: Blaine W
authors.initials: BW
authors.lastname: Phillips
authors.forename: Hannes
authors.initials: H
authors.lastname: Hentze
authors.forename: William L
authors.initials: WL
authors.lastname: Rust
authors.forename: Qi-Ping
authors.initials: QP
authors.lastname: Chen
authors.forename: Hiram
authors.initials: H
authors.lastname: Chipperfield
authors.forename: Ee-Kim
authors.initials: EK
authors.lastname: Tan
authors.forename: Suman
authors.initials: S
authors.lastname: Abraham
authors.forename: Akila
authors.initials: A
authors.lastname: Sadasivam
authors.forename: Poh Loong
authors.initials: PL
authors.lastname: Soong
authors.forename: Siew Tein
authors.initials: ST
authors.lastname: Wang
authors.forename: Ricky
authors.initials: R
authors.lastname: Lim
authors.forename: William
authors.initials: W
authors.lastname: Sun
authors.forename: Alan
authors.initials: A
authors.lastname: Colman
authors.forename: N Ray
authors.initials: NR
authors.lastname: Dunn
mesh_terms: D000818:Animals; D002096:C-Peptide / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D017931:DNA Primers; D053595:Embryonic Stem Cells / Q000166:cytology*; D004797:Enzyme-Linked Immunosorbent Assay; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D018398:Homeodomain Proteins / Q000235:genetics; D006801:Humans; D017403:In Situ Hybridization; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000166:cytology* / Q000502:physiology; D051379:Mice; D016133:Polymerase Chain Reaction; D015534:Trans-Activators / Q000235:genetics