Cells with surface expression of CD133highCD71low are enriched for tripotent colony-forming progenitor cells in the adult murine pancreas

Jin, Liang; Gao, Dan; Feng, Tao; Tremblay, Jacob R.; Ghazalli, Nadiah; Luo, Angela; Rawson, Jeffrey; Quijano, Janine C.; Chai, Jing; Wedeken, Lena; Hsu, Jasper; LeBon, Jeanne M.; Walker, Stephanie; Shih, Hung-Ping; Mahdavi, Alborz; Tirrell, David A.; Riggs, Arthur D.; Ku, Hsun Teresa

doi:10.1016/j.scr.2015.11.015

Cited by 28 publications

(44 citation statements)

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“…Yet, organoids were only formed from CD24 high cells. These cells also had lower expression of CD71 mRNA, which would be consistent with the finding by the Ku group that CD133 high and CD71 low pancreatic cells were enriched in colony-forming units ( Jin et al., 2016 ). More studies are needed to fully characterize these subpopulations.…”

Section: Discussionsupporting

confidence: 90%

“…As previously shown, dissociated single cells from adult mouse pancreas ( Bonner-Weir et al., 2010 , Dorrell et al., 2014 , Huch et al., 2013 , Jin et al., 2016 , Lee et al., 2013 ) and humans ( Lee et al., 2013 ) can be expanded as in vitro organoids in Matrigel. We found about 15% of purified murine duct cells were able to form organoids that can become more than 400 μm in diameter and can be passaged.…”

Section: Discussionmentioning

confidence: 88%

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Heterogeneity of SOX9 and HNF1β in Pancreatic Ducts Is Dynamic

et al. 2018

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SummaryPancreatic duct epithelial cells have been suggested as a source of progenitors for pancreatic growth and regeneration. However, genetic lineage-tracing experiments with pancreatic duct-specific Cre expression have given conflicting results. Using immunofluorescence and flow cytometry, we show heterogeneous expression of both HNF1β and SOX9 in adult human and murine ductal epithelium. Their expression was dynamic and diminished significantly after induced replication. Purified pancreatic duct cells formed organoid structures in 3D culture, and heterogeneity of expression of Hnf1β and Sox9 was maintained even after passaging. Using antibodies against a second cell surface molecule CD51 (human) or CD24 (mouse), we could isolate living subpopulations of duct cells enriched for high or low expression of HNF1β and SOX9. Only the CD24high (Hnfβhigh/Sox9high) subpopulation was able to form organoids.

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Section: Discussionsupporting

confidence: 90%

Section: Discussionmentioning

confidence: 88%

Heterogeneity of SOX9 and HNF1β in Pancreatic Ducts Is Dynamic

et al. 2018

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“…The methods were the same as previously reported [29]. Formalin-fixed, paraffin-embedded cells from murine ES cell-derived cells or postnatal pancreas were processed for sections and staining.…”

Section: Immunohistochemical Stainingmentioning

confidence: 99%

Glucocorticoid Signaling Enhances Expression of Glucose-Sensing Molecules in Immature Pancreatic Beta-Like Cells Derived from Murine Embryonic Stem Cells In Vitro

Ghazalli

Walker

et al. 2018

Stem Cells and Development

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Pluripotent stem cells may serve as an alternative source of beta-like cells for replacement therapy of type 1 diabetes; however, the beta-like cells generated in many differentiation protocols are immature. The maturation of endogenous beta cells involves an increase in insulin expression starting in late gestation and a gradual acquisition of the abilities to sense glucose and secrete insulin by week 2 after birth in mice; however, what molecules regulate these maturation processes are incompletely known. In this study, we aim to identify small molecules that affect immature beta cells. A cell-based assay, using pancreatic beta-like cells derived from murine embryonic stem (ES) cells harboring a transgene containing an insulin 1-promoter driven enhanced green fluorescent protein reporter, was used to screen a compound library (NIH Clinical Collection-003). Cortisone, a glucocorticoid, was among five positive hit compounds. Quantitative reverse transcription-polymerase chain reaction analysis revealed that glucocorticoids enhance the gene expression of not only insulin 1 but also glucose transporter-2 (Glut2; Slc2a2) and glucokinase (Gck), two molecules important for glucose sensing. Mifepristone, a pharmacological inhibitor of glucocorticoid receptor (GR) signaling, reduced the effects of glucocorticoids on Glut2 and Gck expression. The effects of glucocorticoids on ES-derived cells were further validated in immature primary islets. Isolated islets from 1-week-old mice had an increased Glut2 and Gck expression in response to a 4-day treatment of exogenous hydrocortisone in vitro. Gene deletion of GR in beta cells using rat insulin 2 promoter-driven Cre crossed with GR flox/flox mice resulted in a reduced gene expression of Glut2, but not Gck, and an abrogation of insulin secretion when islets were incubated in 0.5 mM d-glucose and stimulated by 17 mM d-glucose in vitro. These results demonstrate that glucocorticoids positively regulate glucose sensors in immature murine beta-like cells.

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“…Discovery of new surface markers and sorting for new subpopulations of pancreatic progenitor-like colonies can help the enrichment of potential cells with high colony-forming efficiency. Some of the specific markers have been discovered, including our previously reported CD71 [ 20 ]. In this study we recorded 7266 mRNA changes in our colonies compared to pancreas.…”

Section: Introductionmentioning

confidence: 99%

Culturing and transcriptome profiling of progenitor-like colonies derived from adult mouse pancreas

Tang

Song

et al. 2017

Stem Cell Res Ther

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BackgroundTransplantation of insulin-producing cells is considered an important diabetes therapy. Many research studies have shown that insulin-producing cells can be derived from the in-vitro cultured pancreatic colonies with self-renewal ability and multilineage potential. Even though these progenitor-like colonies have been prepared from adult pancreas cells, the efficient culture method is hardly established and regulation of the colonies is rarely known. We confirmed previously that single cells acquired from adult mouse pancreas could form cyst-like colonies in a 3D semi-solid system containing Matrigel and methylcellulose. These colonies could be passaged continuously without losing progenitor-like capacity. In the previous culturing system, however, conditioned medium from murine embryonic-stem-cell-derived pancreatic-like cells was used. This unregulated ingredient may reduce repeatability and affect following study. Thus, a new culturing system with certain components needs to be developed.MethodsSingle cell suspension was acquired from adult mouse pancreas and cultured in a Matrigel-based 3D system with epidermal growth factor, Nicotinamide, B27, and Noggin to form ring colonies. Serial-passage assay was performed to evaluate self-renewal ability. Real-time polymerase chain reaction and immunostaining were used to detect the expression of progenitor-related genes. A 2D differentiation method was used to testify the multilineage potency of the colonies. High-throughput sequencing (HTS) of the colonies was performed to profile the differentially expressed genes.ResultsWe developed a 3D culturing system deprived of conditioned medium to propagate those colonies with high proliferative efficiency. HTS of the transcriptome of mRNAs, microRNAs (miRNAs) and long noncoding RNAs (lncRNAs) showed differentially expressed genes compared to the whole pancreas (as control). In mRNAs, several surface marker genes were identified in the colonies. Moreover in noncoding RNAs, miR-21a, miR-31 and miR-155 were upregulated and miR-217, miR-802 and miR-375 were downregulated in colonies along with a number of other miRNAs and lncRNAs.ConclusionsOur results offer an efficient culture system for pancreatic progenitor-like colonies and HTS of the colonies serves as a target resource for following study of in-vitro cultured pancreatic progenitors. These findings should also contribute to our understanding of the transcriptional regulation of these progenitor-like colonies and the mechanisms behind their functions.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-017-0626-y) contains supplementary material, which is available to authorized users.

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Cells with surface expression of CD133highCD71low are enriched for tripotent colony-forming progenitor cells in the adult murine pancreas

Cited by 28 publications

References 45 publications

Heterogeneity of SOX9 and HNF1β in Pancreatic Ducts Is Dynamic

Heterogeneity of SOX9 and HNF1β in Pancreatic Ducts Is Dynamic

Glucocorticoid Signaling Enhances Expression of Glucose-Sensing Molecules in Immature Pancreatic Beta-Like Cells Derived from Murine Embryonic Stem Cells In Vitro

Culturing and transcriptome profiling of progenitor-like colonies derived from adult mouse pancreas

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