GI Tract Development Human Embryonic Stem Or Patrick S

Efficiently differentiate SnapShot: GI Tract Development human embryonic stem or Patrick S. McGrath and James M. Wells Division of Developmental Biology, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH 45229, USA induced pluripotent stem cells to the endoderm Pancreatic lineage allocation Endoderm formation lineage with STEMdiff™ Gastrulation Foregut endoderm Tcf2 (Hnf1β) Use the STEMdiff™ Pancreatic Hnf6 (Onecut1) Mesendoderm Ectoderm Foxa2 (Hnf3β) Progenitor Kit (Catalog #05120) to e7.5 Hlxb9 (Hb9) Hhex generate multipotent PDX-1+/NKX6.1+ Sox17 pancreatic progenitor cells that are Mesoderm Endoderm 200 μm capable of downstream maturation to both endocrine and exocrine cells. Endoderm patterning Anterior Posterior Pancreatic progenitor Pdx1 Use the STEMdiff™ Definitive Nkx2.2 Nkx6.1 Endoderm Kit (Catalog #05110) to Ptf1a (P48) e8.5 Wnt / FGF4 / BMP Hlxb9 (Hb9) generate multipotent definitive endoderm 8–10 somites Poste r cells that are capable of differentiating

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An downstream toward hepatic, intestinal, 300 μm Endocrine progenitor pancreatic and pulmonary cells. Neurog3 Organ speciﬁcation Anterior foregut Posterior Midgut Hindgut NeuroD foregut Ia1 Exocrine progenitor Isl1 Learn more at Pax6 Ptf1a Rfx6 www.stemcell.com/GItract_stemdiff e9.0 Hnf6 ~15 somites Hnf1β STEMCELL Technologies is committed to making sure your research works. As 500 μm Scientists Helping Scientists, we support Organ buds Dorsal pancreas our customers by creating novel products Stomach Duodenum of consistently high quality and by providing Small intestine Thyroid Ventral unparalleled scientiﬁc support. pancreas Pou3f4 (Brn4) Pax4 Nkx2.2 Pax4 ? Liver Large Pax6 MafB Pax6 Biliary intestine MafB Pdx1 e9.5 Lungs Nkx2.2 For legend and references visit ~25 somites Sox2 Cdx www.stemcell.com/wallchart_GItract Pdx1 Hoxa2 Hoxb1 Hoxa3, Hoxb4 Hoxc5 Tbx1 Nkx2.1 Hex1 Ptf1a Hoxc6 Hoxb6 500 μm Hex1 Sox17 Hoxb8,Hoxc9 Hoxb9 Pdx1 PP Sst Cpa Hoxa/d13 Ins Elastase

Proliferation/differentiation Thyroid Lung Biliary tree Stomach Small intestine Colon

>e13.5 α β γ δ ε Exocrine cell Duct Glucagon 50–60 somites Insulin Pancreatic Somatostatin Ghrelin Ptf1a Hnf6 Esophagus Arx Pdx1 polypeptide Pdx1 -Pax4 Gata4 Hnf1β MafB Mnx1 Arx -NKX2.2 Mist1 Sox9 Brn4 Nkx2.2 Amylase Foxa2 Pancreas Nkx6.1 Liver Gall bladder NeuroD MafA

DOCUMENT #27018 | VERSION 1.0.0 SnapShot: GI Tract Development Patrick S. McGrath and James M. Wells Division of Developmental Biology, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH 45229, USA

The endoderm germ layer contributes to the respiratory and gastrointestinal (GI) lineages during development, giving rise to an array of specialized epithelial cell types lining organs, including the thyroid, thymus, lungs, liver, biliary system, pancreas, and intestines. This SnapShot timelines and summarizes key stages following gastrulation, including endoderm patterning, organ specification, and organogenesis. A lineage tree of the developing endocrine pancreas is outlined to further illustrate this process.

Timeline of Endoderm Formation, Patterning, and Organogenesis During development in mice (left), the blastula gives rise to the three germ layers (ectoderm, mesoderm, and definitive endoderm) through the process of gastrulation (middle), which occurs between embryonic day 5 and 7.5 (e5–e7.5). After gastrulation, the two-dimensional sheet of definitive endoderm is patterned along the anterior-posterior (A-P) axis and undergoes morphogenesis to form a three-dimensional gut tube that is surrounded by a primitive mesenchyme (e8.5). A-P patterning of the endoderm occurs through reciprocal signaling with the mesenchyme involving growth factors such as Wnts, Fgfs, and Bmps. At this stage in development, these factors largely act to promote posterior fate and repress anterior fate. The anterior endoderm gives rise to the foregut (thyroid, lungs, esophagus, liver, stomach, pancreas), while the midgut and hindgut give rise to the small and large intestines, respectively. The first evidence of organ specification occurs in the early gut tube by the expression transcription factors that begin to demarcate specific organ domains, including the respiratory tract (Nkx2.1), liver (Hhex), stomach (Sox2 and Pdx1), extrahepatic biliary system (Sox17), pancreas (Pdx1 and Ptf1a), duodenum (Pdx1 and Cdx2), and intestine (Cdx2). The spatially restricted expression of these transcription factors predicts where organs will begin to form starting around e9.5. By e13.5, the organs of the respiratory and GI tracts are formed and undergoing growth and differentiation into specialized lineages.

Pancreatic Lineage Allocation: A Transcription Factor Map Temporal lineage formation of the pancreas involves the expression of unique sets of transcription factors that mark and often direct cell fate decisions (right). All developing cell lineages of the pancreas (acinar, duct, and endocrine) arise from the foregut endoderm, which expresses markers such as Foxa2, Hnf6, and Hlxb9. The pancreatic endoderm becomes specified when the gut tube begins to express Pdx1 and Ptf1a in dorsal and ventral domains of the tube (e8.5–9.0). Morphogenesis of the pancreas initiates with an endodermal thickening (e9.0) and evagination of dorsal and ventral pancreatic buds (e9.5–e10.0) into the surrounding mesenchyme, forming an expanding pool of multipotent pancreatic progenitor cells. The lineage allocation and maturation of specific pancreatic cell subtypes are mediated by a network of signaling pathways and transcription factors. Commitment of progenitor cells to the endocrine lineage occurs following transient expression of Neurog3 and its downstream targets Neurod, Rfx6, and Pax6, whereas exocrine-committed cells express high levels of Ptf1a and carboxypeptidase A (CpA). Allocation of the separate endocrine lineages involves the combinatorial actions of multiple transcription factors. For example, development of mature β cells requires Pdx1, NeuroD, Nx6.1, and MafA. The ductal lineage involves a different set of factors, including Hnf1β and Hnf6. Each pancreatic cell lineage is portrayed with a subset of defining transcription factors throughout development.

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