Download WA27018 GI Tract Development

SnapShot: GI Tract Development
Efficiently differentiate
human embryonic stem or
induced pluripotent stem
cells to the endoderm
lineage with STEMdiff™
Patrick S. McGrath and James M. Wells
Division of Developmental Biology, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH 45229, USA
Pancreatic lineage allocation
Endoderm formation
Gastrulation
Mesendoderm
e7.5
Mesoderm
Foregut endoderm
Tcf2 (Hnf1β)
Hnf6 (Onecut1)
Foxa2 (Hnf3β)
Hlxb9 (Hb9)
Hhex
Sox17
Ectoderm
Use the STEMdiff™ Pancreatic
Progenitor Kit (Catalog #05120) to
generate multipotent PDX-1+/NKX6.1+
pancreatic progenitor cells that are
capable of downstream maturation to
both endocrine and exocrine cells.
Endoderm
200 μm
Endoderm patterning
Posterior
Anterior
e8.5
8–10 somites
Pancreatic progenitor
Pdx1
Nkx2.2
Nkx6.1
Ptf1a (P48)
Hlxb9 (Hb9)
Posterior
Anterior
Wnt / FGF4 / BMP
Wnt-antagonists
300 μm
Organ specification
Anterior foregut
Use the STEMdiff™ Definitive
Endoderm Kit (Catalog #05110) to
generate multipotent definitive endoderm
cells that are capable of differentiating
downstream toward hepatic, intestinal,
pancreatic and pulmonary cells.
Posterior
foregut
Midgut
Hindgut
e9.0
~15 somites
Endocrine progenitor
Neurog3
NeuroD
Ia1
Isl1
Pax6
Rfx6
Exocrine progenitor
Ptf1a
Hnf6
Hnf1β
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500 μm
Organ buds
Dorsal pancreas
Duodenum
Stomach
Thyroid
Liver
Lungs
e9.5
~25 somites
Small intestine
Ventral
pancreas
Large
intestine
Biliary
Sox2
Pou3f4 (Brn4)
Pax6
MafB
Pax4
MafB
Pdx1
Nkx2.2
Nkx2.2
Pdx1
Ins
PP
Pax4
Pax6
Learn more at
www.stemcell.com/GItract_stemdiff
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For legend and references visit
www.stemcell.com/wallchart_GItract
Cdx
Pdx1
Hoxa2
500 μm
Proliferation/differentiation
Hoxb1
Tbx1
Hex1
Thyroid
Hoxa3, Hoxb4
Nkx2.1
Lung
Biliary tree
Hex1
Sox17
Hoxc5
Hoxc6
Ptf1a
Stomach
Small intestine
>e13.5
50–60 somites
Esophagus
Liver
Pancreas
Gall bladder
Hoxb6
Hoxb8,Hoxc9
Hoxb9
Hoxa/d13
Sst
Cpa
Elastase
Colon
α
Glucagon
Arx
MafB
Brn4
β
Insulin
Pdx1
Mnx1
Nkx2.2
Nkx6.1
NeuroD
MafA
γ
δ
ε
Pancreatic Somatostatin Ghrelin
polypeptide
Pdx1
-Pax4
Arx
-NKX2.2
Exocrine cell
Ptf1a
Gata4
Mist1
Amylase
Duct
Hnf6
Hnf1β
Sox9
Foxa2
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.
References
Jørgensen, M.C., Ahnfelt-Rønne, J., Hald, J., Madsen, O.D., Serup, P., and Hecksher-Sørensen, J. (2007). Endocr. Rev. 28, 685–705.
Pagliuca, F.W., and Melton, D.A. (2013). Development 140, 2472–2483.
Pan, F.C., and Wright, C. (2011). Dev. Dyn. 240, 530–565.
Zorn, A.M., and Wells, J.M. (2009). Annu. Rev. Cell Dev. Biol. 25, 221–251.
176.e1 Cell 161, March 26, 2015 ©2015 Elsevier Inc. DOI http://dx.doi.org/10.1016/j.cell.2015.03.014