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Figure 3

Figure 3. Human pluripotent cells are capable of directed differentiation into mature human intestine. From: Generating intestinal tissue from stem cells: potential for research and therapy.

(A) Intestinal organoids can be generated by directed differentiation of human pluripotent cells first into definitive endoderm by low serum and exposure to activin A in vitro. Following culture in WNT3a and FGF4, floating intestinal progenitor spheroids form. When collected and plated at day 0 in the semisolid matrix (Matrigelâ„¢; BD Biosciences, Franklin Lakes, NJ, USA), they grow in size and complexity by 28 days to form epithelium with crypt and villus-like structures (arrowheads). Day 28 organoid epithelium contains all known mature cell types including (B) MUC2-expressing goblet cells, (C) LYZ-expressing Paneth cells and (D) CHGA-expressing enteroendocrine cells.
CHGA: Chromogranin A; LYZ: Lysozyme; MUC: Mucin.

Jonathan C Howell, et al. Regen Med. ;6(6):743-755.
Figure 2

Figure 2. Embryonic development of vertebrate endoderm and intestine. From: Generating intestinal tissue from stem cells: potential for research and therapy.

Following gastrulation of the blastocyst and migration of cells through the primitive streak, the formation of definitive endoderm is induced by exposure to the TGF-b superfamily member Nodal. The endoderm layer eventually forms a primitive gut tube along the anterior–posterior axis that defines proximal and distal gut structures. During gut tube morphogenesis, the influence of various factors establishes patterning of tissue along the anterior–posterior axis such that different domains are able to undergo region-specific organogenesis. During this stage, WNT, FGF and BMP signaling induces hindgut specification and represses the development of foregut tissue. Initial closure of the gut tube occurs at the anterior and posterior ends and proceeds toward the middle until complete. Foregut develops into anterior structures including thyroid, lung, esophagus, stomach, liver, gall bladder and pancreas. Midgut domains develop into small intestine and the proximal large intestine, while hindgut develops into the remainder of the colon.
Adapted with permission from [103,115].

Jonathan C Howell, et al. Regen Med. ;6(6):743-755.
Figure 1

Figure 1. Histology of intestinal layers and crypts. From: Generating intestinal tissue from stem cells: potential for research and therapy.

(A) In cross-section, both small and large intestine contain outer layers of serosa and both longitudinal and circular musculature. The large intestine has large muscular ribbons, called taenia coli, to aid in contraction and peristalsis. Middle layers include submucosa and muscularis mucosa. The innermost layers are the lamina propria and epithelium. In the small intestine, villi project into the intestinal lumen, which expands the surface area available for absorption. Villi are absent in the large intestine. (B) Intestinal crypts contain resident stem cells capable of generating all cell types of the mature epithelium (small intestinal crypt and villus). In the small intestine, Paneth cells provide a crucial microenvironmental niche for stem cells. Paneth cells are largely absent in the large intestine. Following asymmetrical cell division, stem cells give rise to progenitor cells that differentiate into enterocytes (capable of absorption of nutrients and water) hormone secreting enteroendocrine cells, mucin secreting goblet cells and the aforementioned Paneth cells.

Jonathan C Howell, et al. Regen Med. ;6(6):743-755.

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