Epigenetic Induction of Definitive and Pancreatic Endoderm Cell Fate in Human Fibroblasts

Sambathkumar, Rangarajan; Kalo, Eric; Rossom, Rob Van; Faas, Marijke M.; Vos, Paul de; Verfaillie, Catherine M.

doi:10.1155/2016/7654321

Cited by 3 publications

(3 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Before lysis, cells were observed by phase-contrast microscopy, and we noticed that cells treated with TSA, in contrast to AZA treated cells, acquired an elongated morphology (Figure 4A left). The analysis of pluripotency markers such as Oct4 and Nanog [23,24] and differentiation markers such as Nkx6.1 and Pdx1 [25] are in line with the epigenetic pattern of regulation of these genes suggesting the priming of a differentiation program (Figure 4A right).…”

Section: Dna Methylation and Chromatin Acetylation Affect Ink4a And Arf Gene Expression In Escssupporting

confidence: 69%

Pancreatic Progenitor Commitment Is Marked by an Increase in Ink4a/Arf Expression

et al. 2021

View full text Add to dashboard Cite

The identification of the molecular mechanisms controlling early cell fate decisions in mammals is of paramount importance as the ability to determine specific lineage differentiation represents a significant opportunity for new therapies. Pancreatic Progenitor Cells (PPCs) constitute a regenerative reserve essential for the maintenance and regeneration of the pancreas. Besides, PPCs represent an excellent model for understanding pathological pancreatic cellular remodeling. Given the lack of valid markers of early endoderm, the identification of new ones is of fundamental importance. Both products of the Ink4a/Arf locus, in addition to being critical cell-cycle regulators, appear to be involved in several disease pathologies. Moreover, the locus’ expression is epigenetically regulated in ES reprogramming processes, thus constituting the ideal candidates to modulate PPCs homeostasis. In this study, starting from mouse embryonic stem cells (mESCs), we analyzed the early stages of pancreatic commitment. By inducing mESCs commitment to the pancreatic lineage, we observed that both products of the Cdkn2a locus, Ink4a and Arf, mark a naïve pancreatic cellular state that resembled PPC-like specification. Treatment with epi-drugs suggests a role for chromatin remodeling in the CDKN2a (Cycline Dependent Kinase Inhibitor 2A) locus regulation in line with previous observations in other cellular systems. Our data considerably improve the comprehension of pancreatic cellular ontogeny, which could be critical for implementing pluripotent stem cells programming and reprogramming toward pancreatic lineage commitment.

show abstract

Section: Dna Methylation and Chromatin Acetylation Affect Ink4a And Arf Gene Expression In Escssupporting

confidence: 69%

Pancreatic Progenitor Commitment Is Marked by an Increase in Ink4a/Arf Expression

et al. 2021

View full text Add to dashboard Cite

show abstract

“…Inhibition of histone deacetylases drives cells toward differentiation by regulating the expression of differentiation- and pluripotency-associated genes. Many studies demonstrated that TSA enhances cell differentiation [ 4 , 5 , 6 ] by negatively regulating the expression of stemness markers [ 7 ], such as Oct4 [ 8 ], and simultaneously activating differentiation markers, such as Pdx1 [ 9 ]. The molecular mechanism of TSA-mediated regulation of gene activation and repression is very likely related to post-translational modifications of histones.…”

Section: Introductionmentioning

confidence: 99%

Lysines Acetylome and Methylome Profiling of H3 and H4 Histones in Trichostatin A—Treated Stem Cells

Cozzolino

Iacobucci

Monaco

et al. 2021

IJMS

View full text Add to dashboard Cite

Trichostatin A ([R-(E,E)]-7-[4-(dimethylamino) phenyl]-N-hydroxy- 4,6-dimethyl- 7-oxo-2,4-heptadienamide, TSA) affects chromatin state through its potent histone deacetylase inhibitory activity. Interfering with the removal of acetyl groups from lysine residues in histones is one of many epigenetic regulatory processes that control gene expression. Histone deacetylase inhibition drives cells toward the differentiation stage, favoring the activation of specific genes. In this paper, we investigated the effects of TSA on H3 and H4 lysine acetylome and methylome profiling in mice embryonic stem cells (ES14), treated with trichostatin A (TSA) by using a new, untargeted approach, consisting of trypsin-limited proteolysis experiments coupled with MALDI-MS and LC-MS/MS analyses. The method was firstly set up on standard chicken core histones to probe the optimized conditions in terms of enzyme:substrate (E:S) ratio and time of proteolysis and, then, applied to investigate the global variations of the acetylation and methylation state of lysine residues of H3 and H4 histone in the embryonic stem cells (ES14) stimulated by TSA and addressed to differentiation. The proposed strategy was found in its simplicity to be extremely effective in achieving the identification and relative quantification of some of the most significant epigenetic modifications, such as acetylation and lysine methylation. Therefore, we believe that it can be used with equal success in wider studies concerning the characterization of all epigenetic modifications.

show abstract

“…PLGA was used as a cell scaffold to construct the artificial islet tissues, and fibroblasts were used to modify the PLGA membrane to improve the biocompatibility with pancreatic stem cells. Based on related studies on the induction of the differentiation of pancreatic stem cells, [21][22][23][24][25] we used the two-step induction method to induce pancreatic stem cells to differentiate into insulin-producing cells. We constructed a biomodified PLGA membrane, which was a composite cultured with pancreatic stem cells, forming an artificial islet tissue.…”

Section: Introductionmentioning

confidence: 99%

Construction of functional pancreatic artificial islet tissue composed of fibroblast-modified polylactic-co-glycolic acid membrane and pancreatic stem cells

Liu

Tan

et al. 2017

J Biomater Appl

View full text Add to dashboard Cite

Objective To improve the biocompatibility between polylactic- co-glycolic acid membrane and pancreatic stem cells, rat fibroblasts were used to modify the polylactic- co-glycolic acid membrane. Meanwhile, we constructed artificial islet tissue by compound culturing the pancreatic stem cells and the fibroblast-modified polylactic- co-glycolic acid membrane and explored the function of artificial islets in diabetic nude mice. Methods Pancreatic stem cells were cultured on the fibroblast-modified polylactic- co-glycolic acid membrane in dulbecco's modified eagle medium containing activin-A, β-catenin, and exendin-4. The differentiated pancreatic stem cells combined with modified polylactic- co-glycolic acid membrane were implanted subcutaneously in diabetic nude mice. The function of artificial islet tissue was explored by detecting blood levels of glucose and insulin in diabetic nude mice. Moreover, the proliferation and differentiation of pancreatic stem cells on modified polylactic- co-glycolic acid membrane as well as the changes on the tissue structure of artificial islets were investigated by immunofluorescence and haematoxylin and eosin staining. Results The pancreatic stem cells differentiated into islet-like cells and secreted insulin when cultured on fibroblast-modified polylactic- co-glycolic acid membrane. Furthermore, when the artificial islet tissues were implanted into diabetic nude mice, the pancreatic stem cells combined with polylactic- co-glycolic acid membrane modified by fibroblasts proliferated, differentiated, and secreted insulin to reduce blood glucose levels in diabetic nude mice. Conclusion Pancreatic stem cells can be induced to differentiate into islet-like cells in vitro. In vivo, the artificial islet tissue can effectively regulate the blood glucose level in nude mice within a short period. However, as time increased, the structure of the artificial islets was destroyed due to the erosion of blood cells that resulted in the gradual loss of artificial islet function.

show abstract

Epigenetic Induction of Definitive and Pancreatic Endoderm Cell Fate in Human Fibroblasts

Cited by 3 publications

References 34 publications

Pancreatic Progenitor Commitment Is Marked by an Increase in Ink4a/Arf Expression

Pancreatic Progenitor Commitment Is Marked by an Increase in Ink4a/Arf Expression

Lysines Acetylome and Methylome Profiling of H3 and H4 Histones in Trichostatin A—Treated Stem Cells

Construction of functional pancreatic artificial islet tissue composed of fibroblast-modified polylactic-co-glycolic acid membrane and pancreatic stem cells

Contact Info

Product

Resources

About