A Novel Stepwise Differentiation of Functional Pancreatic Exocrine Cells from Embryonic Stem Cells

Shirasawa, Sakiko; Yoshie, Susumu; Yokoyama, Tetsuji; Tomotsune, Daihachiro; Yue, Fengming; Sasaki, Katsunori

doi:10.1089/scd.2010.0185

Cited by 11 publications

(11 citation statements)

References 30 publications

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“…Additional detailed studies will be needed to determine the role of individual differentiating factors in our new method presented here; however, a recent study showed that FGF7 is able to regulate acinar differentiation in mESC [14]. Nonetheless, this protocol was also useful for endocrine differentiation, thus missing a more specific cell lineage approach.…”

Section: Discussionmentioning

confidence: 99%

“…Herein, we aimed to generate exocrine progenitors from mESC following a directed differentiation method. We were interested in enhancing acinar versus endocrine differentiation in vitro , using in the first steps pioneer conditions for definitive endoderm and pancreatic progenitor production [2], [14], [33], [34], [47] and sequentially, a new combination of signals involved in exocrine development.…”

Section: Discussionmentioning

confidence: 99%

“…In this sense, one important aspect missing in many pancreatic differentiation protocols is to assess the extent of selectivity in cell lineage induction. In this regard, other studies have reported the expression of acinar markers from ESC by manipulating several developmental pathways already established for endocrine differentiation or without examining their role on endocrine gene expression [12], [13], [14], [15]. Therefore, progress in the knowledge of how acinar cells are formed during embryogenesis is essential for the improvement of strategies assessing ESC exocrine differentiation.…”

Section: Introductionmentioning

confidence: 99%

“…When this protocol was coupled to high expression of Ptf1a and Rbpjl (designed Ptf1a High and Rbpjl High ), an important rise in the expression of digestive enzymes was observed and cells became responsive to secretagogues. We believe that this new protocol is improved to others in that: i) it favours the generation of exocrine progenitors over the production of the endocrine lineage, ii) it leads to a more mature pattern of the regulatory digestive enzyme expression modules than other reported protocols [11], and iii) generates functional cells in a shorter time [11], [13], [14], [15]. Therefore, this approach might be instrumental for a gain of knowledge of the developmental acinar program and the establishment of in vitro models for the research on pancreatic exocrine disease.…”

Section: Introductionmentioning

confidence: 99%

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Directed Pancreatic Acinar Differentiation of Mouse Embryonic Stem Cells via Embryonic Signalling Molecules and Exocrine Transcription Factors

et al. 2013

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Pluripotent embryonic stem cells (ESC) are a promising cellular system for generating an unlimited source of tissue for the treatment of chronic diseases and valuable in vitro differentiation models for drug testing. Our aim was to direct differentiation of mouse ESC into pancreatic acinar cells, which play key roles in pancreatitis and pancreatic cancer. To that end, ESC were first differentiated as embryoid bodies and sequentially incubated with activin A, inhibitors of Sonic hedgehog (Shh) and bone morphogenetic protein (BMP) pathways, fibroblast growth factors (FGF) and retinoic acid (RA) in order to achieve a stepwise increase in the expression of mRNA transcripts encoding for endodermal and pancreatic progenitor markers. Subsequent plating in Matrigel® and concomitant modulation of FGF, glucocorticoid, and folllistatin signalling pathways involved in exocrine differentiation resulted in a significant increase of mRNAs encoding secretory enzymes and in the number of cells co-expressing their protein products. Also, pancreatic endocrine marker expression was down-regulated and accompanied by a significant reduction in the number of hormone-expressing cells with a limited presence of hepatic marker expressing-cells. These findings suggest a selective activation of the acinar differentiation program. The newly differentiated cells were able to release α-amylase and this feature was greatly improved by lentiviral-mediated expression of Rbpjl and Ptf1a, two transcription factors involved in the maximal production of digestive enzymes. This study provides a novel method to produce functional pancreatic exocrine cells from ESC.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Directed Pancreatic Acinar Differentiation of Mouse Embryonic Stem Cells via Embryonic Signalling Molecules and Exocrine Transcription Factors

et al. 2013

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“…FGF7 отвечает в основном за пролиферацию эпителиальных клеток, но в то же время влияет на гены, регулирующие программу развития поджелудочной железы, такие как Pdx1, Nkx6.1, Hnf6, Ptf1a [39]. FGF10 способен удерживать клетки в недифференцированном состоянии и активировать мастер-ген Sox9 и несколько других важных сигнальных путей [40].…”

Section: Fgfr2unclassified

Role of fibroblast growth factors in pancreatic cancer

Gnatenko

Копанцев

2016

BIOMED KHIM

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Факторы роста фибробластов (FGF) -сигнальные молекулы c широким спектром действия, вовлечённые во многие клеточные процессы. FGF играют важную роль не только в органогенезе, но и в канцерогенезе поджелудочной железы. Они относятся к тем факторам, посредством которых происходит потенцирующее взаимодействие между опухолью и окружающей её стромой -ключевым компонентом развития рака поджелудочной железы. Нарушения сигнального пути FGF/FGFR -комплексный процесс, в котором важную роль играют тип и изоформа рецепторов, регулирующих ремоделирующий эффект FGF и последующую активацию данными факторами пролиферации клеток рака поджелудочной железы. Факторы FGF и их рецепторы рассматриваются как потенциальные специфические маркеры и возможные мишени для терапии рака поджелудочной железы.Ключевые слова: ростовые факторы, рак, поджелудочная железа, мастер-гены

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FGF7 and cell density are required for final differentiation of pancreatic amylase-positive cells from human ES cells

et al. 2013

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The major molecular signals of pancreatic exocrine development are largely unknown. We examine the role of fibroblast growth factor 7 (FGF7) in the final induction of pancreatic amylase-containing exocrine cells from induced-pancreatic progenitor cells derived from human embryonic stem (hES) cells. Our protocol consisted in three steps: Step I, differentiation of definitive endoderm (DE) by activin A treatment of hES cell colonies; Step II, differentiation of pancreatic progenitor cells by re-plating of the cells of Step I onto 24-well plates at high density and stimulation with all-trans retinoic acid; Step III, differentiation of pancreatic exocrine cells with a combination of FGF7, glucagon-like peptide 1 and nicotinamide. The expression levels of pancreatic endodermal markers such as Foxa2, Sox17 and gut tube endoderm marker HNF1β were up-regulated in both Step I and II. Moreover, in Step III, the induced cells expressed pancreatic markers such as amylase, carboxypeptidase A and chymotrypsinogen B, which were similar to those in normal human pancreas. From day 8 in Step III, cells immunohistochemically positive for amylase and for carboxypeptidase A, a pancreatic exocrine cell product, were induced by FGF7. Pancreatic progenitor Pdx1-positive cells were localized in proximity to the amylase-positive cells. In the absence of FGF7, few amylase-positive cells were identified. Thus, our three-step culture protocol for human ES cells effectively induces the differentiation of amylase- and carboxypeptidase-A-containing pancreatic exocrine cells.

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A Novel Stepwise Differentiation of Functional Pancreatic Exocrine Cells from Embryonic Stem Cells

Cited by 11 publications

References 30 publications

Directed Pancreatic Acinar Differentiation of Mouse Embryonic Stem Cells via Embryonic Signalling Molecules and Exocrine Transcription Factors

Directed Pancreatic Acinar Differentiation of Mouse Embryonic Stem Cells via Embryonic Signalling Molecules and Exocrine Transcription Factors

Role of fibroblast growth factors in pancreatic cancer

FGF7 and cell density are required for final differentiation of pancreatic amylase-positive cells from human ES cells

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