Differentiation of Adult Hepatic Stem-Like Cells into Pancreatic Endocrine Cells

To apply cell transplantation for treatment of diabetes mellitus, a sufficient number of β-cell sources are required. In the present study, we examined whether an epithelial cell line obtained from normal adult rat liver, namely hepatic stem-like (HSL) cells, which can be converted to both hepatocytes and billiary epithelial cells, could be a potential β-cell source. The growth speed of HSL cells was rapid and these cells were easily expanded in vitro. Bipotential hepatic stem cells, HSL cells, also expressed PGP9.5, which is expressed in neurons, β-cells, and progenitor cells of the pancreatic endocrine cells as well. Sodium butyrate induced morphological changes in HSL cells and converted them into flattened cells with large cytoplasm. When HSL cells were incubated with a combination of 5 mM sodium butyrate and 1 nM betacellulin, most of the cells were converted into morphologically neuron-like cells. RT-PCR analysis revealed that a series of transcriptional factors involved in differentiation of pancreatic endocrine cells was induced by the treatment with sodium butyrate and betacellulin. mRNAs for insulin, pancreatic polypeptide, and somatostatin were also observed. Immunoreactive pancreatic polypeptide, somatostatin, and insulin were detected in sodium butyrate and betacellulin-treated HSL cells. In conclusion, HSL cells obtained from adult normal liver also have the potential to differentiate into pancreatic endocrine cells in vitro. HSL cells may be one of the potential β-cell sources for cell transplant therapy for insulin-dependent diabetes.

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Human Pluripotent Stem Cells: A Unique Tool for Toxicity Testing in Pancreatic Progenitor and Endocrine Cells

Frontiers in Endocrinology ◽

10.3389/fendo.2020.604998 ◽

2021 ◽

Vol 11 ◽

Author(s):

Erin M. MacFarlane ◽

Jennifer E. Bruin

Keyword(s):

High Throughput ◽

Endocrine Cells ◽

Toxicity Testing ◽

Diabetes Incidence ◽

Clear Understanding ◽

Target Tissue ◽

Β Cell ◽

Obesity And Diabetes ◽

Pancreatic Endocrine Cells

Diabetes prevalence is increasing worldwide, and epidemiological studies report an association between diabetes incidence and environmental pollutant exposure. There are >84,000 chemicals in commerce, many of which are released into the environment without a clear understanding of potential adverse health consequences. While in vivo rodent studies remain an important tool for testing chemical toxicity systemically, we urgently need high-throughput screening platforms in biologically relevant models to efficiently prioritize chemicals for in depth toxicity analysis. Given the increasing global burden of obesity and diabetes, identifying chemicals that disrupt metabolism should be a high priority. Pancreatic endocrine cells are key regulators of systemic metabolism, yet often overlooked as a target tissue in toxicology studies. Immortalized β-cell lines and primary human, porcine, and rodent islets are widely used for studying the endocrine pancreas in vitro, but each have important limitations in terms of scalability, lifespan, and/or biological relevance. Human pluripotent stem cell (hPSC) culture is a powerful tool for in vitro toxicity testing that addresses many of the limitations with other β-cell models. Current in vitro differentiation protocols can efficiently generate glucose-responsive insulin-secreting β-like cells that are not fully mature, but still valuable for high-throughput toxicity screening in vitro. Furthermore, hPSCs can be applied as a model of developing pancreatic endocrine cells to screen for chemicals that influence endocrine cell formation during critical windows of differentiation. Given their versatility, we recommend using hPSCs to identify potential β-cell toxins, which can then be prioritized as chemicals of concern for metabolic disruption.

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The distribution of endocrine cell progenitors in the gut of chick embryos

Development ◽

10.1242/dev.82.1.131 ◽

1984 ◽

Vol 82 (1) ◽

pp. 131-145

Author(s):

B. B. Rawdon ◽

Beverley Kramer ◽

Ann Andrew

Keyword(s):

Gastrointestinal Tract ◽

Progenitor Cells ◽

Digestive Tract ◽

Pancreatic Polypeptide ◽

Endocrine Cell ◽

Endocrine Cells ◽

Chick Embryos ◽

Intact Embryo ◽

Pancreatic Endocrine Cells ◽

Gut Endocrine Cells

The aim of this experiment was to find out whether or not, at early stages of development, progenitors of the various types of gut endocrine cells are localized to one or more specific regions of the gastrointestinal tract. Transverse strips of blastoderm two to four somites in length were excised between the levels of somites 5 and 27 in chick embryos at 5- to 24-somite stages and were cultured as chorioallantoic grafts. The distribution of endocrine cells in the grafts revealed confined localization of progenitor cells only in the case of insulinimmunoreactive cells. Theprogenitors of cells with somatostatin-, pancreatic polypeptide-, glucagon-, secretin-, gastrin/CCK-, motilin-, neurotensin- and serotonin-like immunoreactivity were distributed along the length of the presumptive gut at the time of explantation; indeed, in many cases they were more widespread than are their differentiated progeny in normal gut of the same age. This finding indicates that conditions in grafts must differ from those that operate in the intact embryo. Also it may explain the occurrence of ectopic gut or pancreatic endocrine cells in tumours of the digestive tract.

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REST is a major negative regulator of endocrine differentiation during pancreas organogenesis

10.1101/2021.03.17.435876 ◽

2021 ◽

Author(s):

Meritxell Rovira ◽

Goutham Atla ◽

Miguel Angel Maestro ◽

Vane Grau ◽

Javier García-Hurtado ◽

...

Keyword(s):

Endocrine Cells ◽

Negative Regulator ◽

Β Cell ◽

Knock Out ◽

Endocrine Differentiation ◽

Conditional Allele ◽

Embryonic Pancreas ◽

Pancreatic Endocrine Cells ◽

Knock Out Mice

SUMMARYUnderstanding genomic regulatory mechanisms of pancreas differentiation is relevant to the pathophysiology of diabetes mellitus, and to the development of replacement therapies. Numerous transcription factors promote β cell differentiation, although less is known about negative regulators. Earlier epigenomic studies suggested that the transcriptional repressor REST could be a suppressor of endocrine gene programs in the embryonic pancreas. However, pancreaticRestknock-out mice failed to show increased numbers of endocrine cells, suggesting that REST is not a major regulator of endocrine differentiation. Using a different conditional allele that enables profound REST inactivation, we now observe a marked increase in the formation of pancreatic endocrine cells. REST inhibition also promoted endocrinogenesis in zebrafish and mouse early postnatal ducts, and induced β-cell specific genes in human adult duct-derived organoids. Finally, we define REST genomic programs that suppress pancreatic endocrine differentiation. These results establish a crucial role of REST as a negative regulator of pancreatic endocrine differentiation.

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ASCR-007 Chronological change of gene expression during the transdifferentiation from human amniotic epithelial cells to pancreatic endocrine cells in vitro

Reproductive BioMedicine Online ◽

10.1016/s1472-6483(10)61542-8 ◽

2008 ◽

Vol 16 ◽

pp. S-31-S-32

Author(s):

L Peng ◽

J Wang ◽

GX Lu

Keyword(s):

Gene Expression ◽

Epithelial Cells ◽

Endocrine Cells ◽

Chronological Change ◽

Amniotic Epithelial Cells ◽

Pancreatic Endocrine Cells ◽

Human Amniotic Epithelial Cells

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Expansion and conversion of human pancreatic ductal cells into insulin-secreting endocrine cells

eLife ◽

10.7554/elife.00940 ◽

2013 ◽

Vol 2 ◽

Cited By ~ 88

Author(s):

Jonghyeob Lee ◽

Takuya Sugiyama ◽

Yinghua Liu ◽

Jing Wang ◽

Xueying Gu ◽

...

Keyword(s):

Endocrine Cells ◽

General Strategy ◽

In Vitro Production ◽

Β Cell ◽

Ductal Cells ◽

Adult Human ◽

Pancreatic Ductal Cells ◽

Pancreatic Cells ◽

Vitro Production

Pancreatic islet β-cell insufficiency underlies pathogenesis of diabetes mellitus; thus, functional β-cell replacement from renewable sources is the focus of intensive worldwide effort. However, in vitro production of progeny that secrete insulin in response to physiological cues from primary human cells has proven elusive. Here we describe fractionation, expansion and conversion of primary adult human pancreatic ductal cells into progeny resembling native β-cells. FACS-sorted adult human ductal cells clonally expanded as spheres in culture, while retaining ductal characteristics. Expression of the cardinal islet developmental regulators Neurog3, MafA, Pdx1 and Pax6 converted exocrine duct cells into endocrine progeny with hallmark β-cell properties, including the ability to synthesize, process and store insulin, and secrete it in response to glucose or other depolarizing stimuli. These studies provide evidence that genetic reprogramming of expandable human pancreatic cells with defined factors may serve as a general strategy for islet replacement in diabetes.

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Immunocytochemical study of the distribuition of endocrine cells in the pancreas of the Brazilian sparrow species Zonotrichia Capensis Subtorquata (Swaison, 1837)

Brazilian Journal of Biology ◽

10.1590/s1519-69842007000400021 ◽

2007 ◽

Vol 67 (4) ◽

pp. 735-740 ◽

Cited By ~ 8

Author(s):

AA Nascimento ◽

A Sales ◽

TRD Cardoso ◽

NL Pinheiro ◽

RMM Mendes

Keyword(s):

B Cells ◽

Pancreatic Islets ◽

Pancreatic Polypeptide ◽

Endocrine Cells ◽

Central Place ◽

Immunocytochemical Study ◽

Zonotrichia Capensis ◽

Specific Antisera ◽

Pancreatic Endocrine Cells ◽

Avian Pancreatic Polypeptide

In the present study, we investigated types of pancreatic endocrine cells and its respective peptides in the Brazilian sparrow species using immunocytochemistry. The use of polyclonal specific antisera for somatostatin, glucagon, avian pancreatic polypeptide (APP), YY polypeptide (PYY) and insulin, revealed a diversified distribution in the pancreas. All these types of immunoreactive cells were observed in the pancreas with different amounts. Insulin- Immunoreactive cells to (B cells) were most numerous, preferably occupying the central place in the pancreatic islets. Somatostatin, PPA, PYY and glucagon immunoreactive cells occurred in a lower frequency in the periphery of pancreatic islets.

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In Vitro Generation of Pancreatic Endocrine Cells from Human Adult Fibroblast-Like Limbal Stem Cells

Cell Transplantation ◽

10.3727/096368911x580635 ◽

2012 ◽

Vol 21 (1) ◽

pp. 73-90 ◽

Cited By ~ 10

Author(s):

Angela Criscimanna ◽

Giovanni Zito ◽

Annalisa Taddeo ◽

Pierina Richiusa ◽

Maria Pitrone ◽

...

Keyword(s):

Stem Cells ◽

Endocrine Cells ◽

Limbal Stem Cells ◽

Human Adult ◽

Pancreatic Endocrine Cells

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Development and Characteristics of Pancreatic Epsilon Cells

International Journal of Molecular Sciences ◽

10.3390/ijms20081867 ◽

2019 ◽

Vol 20 (8) ◽

pp. 1867 ◽

Cited By ~ 5

Author(s):

Naoaki Sakata ◽

Gumpei Yoshimatsu ◽

Shohta Kodama

Keyword(s):

Transcription Factors ◽

Blood Glucose ◽

Endocrine Cells ◽

Current Evidence ◽

Β Cells ◽

Β Cell ◽

Glucose Levels ◽

Blood Glucose Levels ◽

Ghrelin Gene ◽

Pancreatic Endocrine Cells

Pancreatic endocrine cells expressing the ghrelin gene and producing the ghrelin hormone were first identified in 2002. These cells, named ε cells, were recognized as the fifth type of endocrine cells. Differentiation of ε cells is induced by various transcription factors, including Nk2 homeobox 2, paired box proteins Pax-4 and Pax6, and the aristaless-related homeobox. Ghrelin is generally considered to be a “hunger hormone” that stimulates the appetite and is produced mainly by the stomach. Although the population of ε cells is small in adults, they play important roles in regulating other endocrine cells, especially β cells, by releasing ghrelin. However, the roles of ghrelin in β cells are complex. Ghrelin contributes to increased blood glucose levels by suppressing insulin release from β cells and is also involved in the growth and proliferation of β cells and the prevention of β cell apoptosis. Despite increasing evidence and clarification of the mechanisms of ε cells over the last 20 years, many questions remain to be answered. In this review, we present the current evidence for the participation of ε cells in differentiation and clarify their characteristics by focusing on the roles of ghrelin.

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Mitchell-Riley syndrome iPSCs exhibit reduced pancreatic endoderm differentiation due to a mutation in RFX6

Development ◽

10.1242/dev.194878 ◽

2020 ◽

Vol 147 (21) ◽

pp. dev194878

Author(s):

Jamie Trott ◽

Yunus Alpagu ◽

Ee Kim Tan ◽

Mohammad Shboul ◽

Yousif Dawood ◽

...

Keyword(s):

Endocrine Cells ◽

Neonatal Diabetes ◽

Pancreas Head ◽

Recessive Mutations ◽

Pancreatic Endocrine Cells ◽

Pancreas Body ◽

Pancreatic Hypoplasia

ABSTRACTMitchell-Riley syndrome (MRS) is caused by recessive mutations in the regulatory factor X6 gene (RFX6) and is characterised by pancreatic hypoplasia and neonatal diabetes. To determine why individuals with MRS specifically lack pancreatic endocrine cells, we micro-CT imaged a 12-week-old foetus homozygous for the nonsense mutation RFX6 c.1129C>T, which revealed loss of the pancreas body and tail. From this foetus, we derived iPSCs and show that differentiation of these cells in vitro proceeds normally until generation of pancreatic endoderm, which is significantly reduced. We additionally generated an RFX6HA reporter allele by gene targeting in wild-type H9 cells to precisely define RFX6 expression and in parallel performed in situ hybridisation for RFX6 in the dorsal pancreatic bud of a Carnegie stage 14 human embryo. Both in vitro and in vivo, we find that RFX6 specifically labels a subset of PDX1-expressing pancreatic endoderm. In summary, RFX6 is essential for efficient differentiation of pancreatic endoderm, and its absence in individuals with MRS specifically impairs formation of endocrine cells of the pancreas head and tail.

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