Osteopontin is a novel marker of pancreatic ductal tissues and of undifferentiated pancreatic precursors in mice

Kılıç, Gamze; Wang, Junfeng; Sosa–Pineda, Beatriz

doi:10.1002/dvdy.20729

Cited by 31 publications

(27 citation statements)

References 34 publications

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“…To confirm protein expression, 3-wk-old Ring colonies were handpicked, pooled, fixed, and analyzed by whole-mount or frozensection immunofluorescent staining and confocal imaging. Consistent with the microfluidic qRT-PCR analysis, ductal protein marker expression was present in all colonies examined, and each colony was positive for ductal proteins, Sox9/EGFP, Mucin1, or Osteopontin1 (Spp1) (52) (Fig. 1E).…”

Section: Sox9/egfpsupporting

confidence: 78%

Colony-forming cells in the adult mouse pancreas are expandable in Matrigel and form endocrine/acinar colonies in laminin hydrogel

Jin

Feng

Shih

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

183

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The study of hematopoietic colony-forming units using semisolid culture media has greatly advanced the knowledge of hematopoiesis. Here we report that similar methods can be used to study pancreatic colony-forming units. We have developed two pancreatic colony assays that enable quantitative and functional analyses of progenitor-like cells isolated from dissociated adult (2-4 mo old) murine pancreas. We find that a methylcellulose-based semisolid medium containing Matrigel allows growth of duct-like "Ring/ Dense" colonies from a rare (∼1%) population of total pancreatic single cells. With the addition of roof plate-specific spondin 1, a wingless-int agonist, Ring/Dense colony-forming cells can be expanded more than 100,000-fold when serially dissociated and replated in the presence of Matrigel. When cells grown in Matrigel are then transferred to a Matrigel-free semisolid medium with a unique laminin-based hydrogel, some cells grow and differentiate into another type of colony, which we name "Endocrine/Acinar." These Endocrine/Acinar colonies are comprised mostly of endocrine-and acinar-like cells, as ascertained by RNA expression analysis, immunohistochemistry, and electron microscopy. Most Endocrine/Acinar colonies contain beta-like cells that secrete insulin/C-peptide in response to D-glucose and theophylline. These results demonstrate robust self-renewal and differentiation of adult Ring/Dense colony-forming units in vitro and suggest an approach to producing beta-like cells for cell replacement of type 1 diabetes. The methods described, which include microfluidic expression analysis of single cells and colonies, should also advance study of pancreas development and pancreatic progenitor cells. extracellular matrix proteins | Sry-related HMG box (Sox) 9 | Promonin 1 (CD133) | neurogenin 3 | dickkopf1 (Dkk1)

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Section: Sox9/egfpsupporting

confidence: 78%

Colony-forming cells in the adult mouse pancreas are expandable in Matrigel and form endocrine/acinar colonies in laminin hydrogel

Jin

Feng

Shih

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

183

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“…S4D in the supplementary material), whereas OPN also marks endocrine cells (Kilic et al, 2006; data not shown). Our findings show that, especially during development, presumed ductal markers are not expressed in identical cellular domains, implying that these domains could differ in their developmental potential.…”

Section: Research Articlementioning

confidence: 80%

Sox9+ ductal cells are multipotent progenitors throughout development but do not produce new endocrine cells in the normal or injured adult pancreas

et al. 2011

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SUMMARYOne major unresolved question in the field of pancreas biology is whether ductal cells have the ability to generate insulinproducing b-cells. Conclusive examination of this question has been limited by the lack of appropriate tools to efficiently and specifically label ductal cells in vivo. We generated Sox9CreER T2 mice, which, during adulthood, allow for labeling of an average of 70% of pancreatic ductal cells, including terminal duct/centroacinar cells. Fate-mapping studies of the Sox9 + domain revealed endocrine and acinar cell neogenesis from Sox9 + cells throughout embryogenesis. Very small numbers of non-b endocrine cells continue to arise from Sox9 + cells in early postnatal life, but no endocrine or acinar cell neogenesis from Sox9 + cells occurs during adulthood. In the adult pancreas, pancreatic injury by partial duct ligation (PDL) has been suggested to induce b-cell regeneration from a transient Ngn3 + endocrine progenitor cell population. Here, we identify ductal cells as a cell of origin for PDL-induced Ngn3 + cells, but fail to observe b-cell neogenesis from duct-derived cells. Therefore, although PDL leads to activation of Ngn3 expression in ducts, PDL does not induce appropriate cues to allow for completion of the entire b-cell neogenesis program. In conclusion, although endocrine cells arise from the Sox9 + ductal domain throughout embryogenesis and the early postnatal period, Sox9 + ductal cells of the adult pancreas no longer give rise to endocrine cells under both normal conditions and in response to PDL.

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“…Overexpression was performed by adenoviral transduction of HNF6 (Ad-HNF6) or Sox9 (Ad-Sox9), with GFP-expressing adenovirus (Ad-GFP) as a control. In the acinar 266-6 cell line, expression of HNF6 (but not of GFP or Sox9) induced Sox9, CK19 and osteopontin30 which, in the pancreas, have an expression restricted to the ductal cells, and repressed the acinar markers Mist1, Ptf1a, amylase, CPA and elastase (figure 2A). HNF1β was weakly induced by HNF6 (figure 2A).…”

Section: Resultsmentioning

confidence: 99%

Role of the ductal transcription factors HNF6 and Sox9 in pancreatic acinar-to-ductal metaplasia

Prévôt

Simion

Grimont

et al. 2012

Gut

111

103

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Objective Growing evidence suggests that a phenotypic switch converting pancreatic acinar cells to duct-like cells can lead to pancreatic intraepithelial neoplasia (PanIN) and eventually to invasive pancreatic ductal adenocarcinoma. Histologically, the onset of this switch is characterised by the co-expression of acinar and ductal markers in acini, a lesion called acinar-to-ductal metaplasia (ADM). Transcriptional regulators required to initiate ADM still remain unknown, yet need to be identified to characterise the regulatory networks that drive ADM. Here we investigate the role of the ductal transcription factors Hepatocyte Nuclear Factor 6 (HNF6, also known as Onecut1)and SRY-related HMG box factor 9 (Sox9) in ADM. Design Expression of HNF6 and Sox9 is measured by immunostaining in normal and diseased human pancreas. The function of the factors is tested in cultured cells and in mouse models of ADM by a combination of gain- and loss-of-function experiments. Results Expression of HNF6 and Sox9 is ectopically induced in acinar cells in human ADM, as well as in mouse models of ADM. We show that these factors are required for repression of acinar genes, for modulation of ADM-associated changes in cell polarity, and for activation of ductal genes in metaplastic acinar cells. Conclusions HNF6 and Sox9 are new biomarkers of ADM and constitute candidate targets for preventive therapy in cases when ADM may lead to cancer. Our work also highlights that ectopic activation of transcription factors may underlie metaplastic processes occurring in other organs.

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Osteopontin is a novel marker of pancreatic ductal tissues and of undifferentiated pancreatic precursors in mice

Cited by 31 publications

References 34 publications

Colony-forming cells in the adult mouse pancreas are expandable in Matrigel and form endocrine/acinar colonies in laminin hydrogel

Colony-forming cells in the adult mouse pancreas are expandable in Matrigel and form endocrine/acinar colonies in laminin hydrogel

Sox9+ ductal cells are multipotent progenitors throughout development but do not produce new endocrine cells in the normal or injured adult pancreas

Role of the ductal transcription factors HNF6 and Sox9 in pancreatic acinar-to-ductal metaplasia

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