Reconstituting pancreas development from purified progenitor cells reveals genes essential for islet differentiation

Sugiyama, Takuya; Benitez, Cecil M.; Ghodasara, Amar; Liu, Lucy; McLean, Graeme; Lee, Jonghyeob; Blauwkamp, Timothy A.; Nusse, Roeland; Wright, Christopher V.E.; Gu, Guoqiang; Kim, Seung K.

doi:10.1073/pnas.1304507110

Cited by 65 publications

(76 citation statements)

References 50 publications

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“…In future experiments we hope to combine the clonal culture of pancreas progenitors with cell sorting to analyze defined progenitor cell populations at different stages of development. They share similarities with the spheres recently published by Sugiyama et al (Sugiyama et al, 2013). By using methods such as multiplexed single-cell gene expression analysis one should be able to provide a paradigm to test the response of single cells to the perturbation of a specific signaling pathway or to changes in extracellular components.…”

Section: Research Articlementioning

confidence: 65%

Artificial three-dimensional niches deconstruct pancreas developmentin vitro

et al. 2013

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SUMMARYIn the context of a cellular therapy for diabetes, methods for pancreatic progenitor expansion and subsequent differentiation into insulin-producing beta cells would be extremely valuable. Here we establish three-dimensional culture conditions in Matrigel that enable the efficient expansion of dissociated mouse embryonic pancreatic progenitors. By manipulating the medium composition we generate either hollow spheres, which are mainly composed of pancreatic progenitors, or complex organoids that spontaneously undergo pancreatic morphogenesis and differentiation. The in vitro maintenance and expansion of pancreatic progenitors require active Notch and FGF signaling, thus recapitulating in vivo niche signaling interactions. Our experiments reveal new aspects of pancreas development, such as a community effect by which small groups of cells better maintain progenitor properties and expand more efficiently than isolated cells, as well as the requirement for three-dimensionality. Finally, growth conditions in chemically defined biomaterials pave the way for testing the biophysical and biochemical properties of the niche that sustains pancreatic progenitors.

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Section: Research Articlementioning

confidence: 65%

Artificial three-dimensional niches deconstruct pancreas developmentin vitro

et al. 2013

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“…(1,41). Prdm16 was expressed at the highest levels in iBAT, followed by ingWAT and with some expression in eWAT (Fig.…”

Section: Resultsmentioning

confidence: 99%

A stringent validation of mouse adipose tissue identity markers

Jong

Larsson

Cannon

et al. 2015

American Journal of Physiology-Endocrinology and Metabolism

241

212

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“…Other existing culture assays for epithelial cells employ Matrigel but not methylcellulose. In those assays, epithelial cells of gut [29], liver [30], or pancreas [31,32] are mixed in high concentrations ( > 60% vol/vol) of Matrigel, which is required to render a 3D culture environment. Subsequently, liquid media are layered over on top of the Matrigel.…”

Section: Discussionmentioning

confidence: 99%

In Vitro Multilineage Differentiation and Self-Renewal of Single Pancreatic Colony-Forming Cells from Adult C57Bl/6 Mice

Jin

Feng

Zerda

et al. 2014

Stem Cells and Development

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In a previous study we established colony assays suitable for studying murine adult (2-4 months) pancreatic progenitor cells plated in semisolid media containing methylcellulose and extracellular matrix proteins. Using these assays, we found robust in vitro progenitor cell activities (multilineage differentiation and self-renewal) from pancreatic cells of adult mice in the CD-1 outbred background. However, it was not clear whether progenitor cell activities can be detected from inbred mice, a preferred mouse model for various genetic studies. It was also not clear whether a single cell is sufficient to self-renew. Here, we show that fluorescent activated cell sorting pancreatic CD133+ but not CD133 -cells from adult C57Bl/6 inbred mice are enriched for progenitor cells that self-renew and give rise to multilineage colonies in vitro. The number of cells in a colony is in proportion to its diameter. Around 60% of single handpicked 3-week-old colonies express trilineage markers, indicating most progenitors are tripotent for ductal, acinar, and endocrine lineage differentiation. Approximately 80% of primary (freshly sorted) colony-forming progenitor cells are capable of giving rise to secondary progenitors in vitro, indicating that a majority of the primary progenitors self-renew. A single cell is sufficient for self-renewal and a Wnt agonist, R-Spondin1, enhances the number of secondary progenitors from the primary progenitors. Together, our pancreatic colony assays allow quantitative analyses of progenitors at a single-cell level from inbred mice. These assays will be useful for elucidating in vitro mechanisms of pancreatic progenitor cell biology.

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Reconstituting pancreas development from purified progenitor cells reveals genes essential for islet differentiation

Cited by 65 publications

References 50 publications

Artificial three-dimensional niches deconstruct pancreas developmentin vitro

Artificial three-dimensional niches deconstruct pancreas developmentin vitro

A stringent validation of mouse adipose tissue identity markers

In Vitro Multilineage Differentiation and Self-Renewal of Single Pancreatic Colony-Forming Cells from Adult C57Bl/6 Mice

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