MicroRNA expression profile during adipogenic differentiation in mouse embryonic stem cells

2011 ◽  
Vol 43 (10) ◽  
pp. 611-620 ◽  
Author(s):  
Julia M. Knelangen ◽  
Mark B. van der Hoek ◽  
Wee-Ching Kong ◽  
Julie A. Owens ◽  
Bernd Fischer ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Cell Fate ◽  
Adipogenic Differentiation ◽  
Embryonic Stem Cell Line ◽  
Embryonic Stem ◽  
Mouse Embryonic Stem Cell ◽  
Specific Cell ◽  
Cell Fate Determination ◽  
Fate Determination

Pluripotent embryonic stem cells (ESC) have the potential to differentiate into any cell type of the three germ layers. Differentiation processes depend on genetic and epigenetic factors. The guidance of cell fate determination by microRNAs (miRs) seems important for embryonic development and cell lineage decisions. MiRs are short, single-stranded, noncoding RNA molecules that regulate through posttranscriptional modulation, a subset of target genes involved in cell differentiation and specific cell function. We have used microarray profiling of miRs in the mouse embryonic stem cell line CGR8. Comparison of the miR profiles of undifferentiated stem cells with mesodermal progenitors cells ( day 5), preadipocytes ( day 10), and adipocytes ( day 21) showed that the expression level of 129 miRs changed (twofold) during adipogenic differentiation. We identified 10 clusters of differentially expressed miRs, which contain putative markers and regulators of mesodermal differentiation and cell fate determination into adipocytes. Notably, the adipocyte-specific miRs 143 and 103 were upregulated from day 10 onward. We have therefore demonstrated and characterized the dynamic profile of miR expression during murine adipogenic differentiation in vitro, including the initial differentiation from ESC via mesenchymal progenitors up to adipocytes. Our findings and experimental approach provide a suitable system to directly interrogate the role of miRs during adipogenic differentiation of embryonic stem cells.

scholarly journals Nicotinamide promotes pancreatic differentiation through the dual inhibition of CK1 and ROCK kinases in human embryonic stem cells

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Yumeng Zhang ◽  
Jiaqi Xu ◽  
Zhili Ren ◽  
Ya Meng ◽  
Weiwei Liu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Fate ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Cell Fate Determination ◽  
Rna Seq ◽  
Pancreatic Cell ◽  
Pancreatic Progenitor ◽  
Pancreatic Progenitors ◽  
Rock Inhibition

Abstract Background Vitamin B3 (nicotinamide) plays important roles in metabolism as well as in SIRT and PARP pathways. It is also recently reported as a novel kinase inhibitor with multiple targets. Nicotinamide promotes pancreatic cell differentiation from human embryonic stem cells (hESCs). However, its molecular mechanism is still unclear. In order to understand the molecular mechanism involved in pancreatic cell fate determination, we analyzed the downstream pathways of nicotinamide in the derivation of NKX6.1+ pancreatic progenitors from hESCs. Methods We applied downstream modulators of nicotinamide during the induction from posterior foregut to pancreatic progenitors, including niacin, PARP inhibitor, SIRT inhibitor, CK1 inhibitor and ROCK inhibitor. The impact of those treatments was evaluated by quantitative real-time PCR, flow cytometry and immunostaining of pancreatic markers. Furthermore, CK1 isoforms were knocked down to validate CK1 function in the induction of pancreatic progenitors. Finally, RNA-seq was used to demonstrate pancreatic induction on the transcriptomic level. Results First, we demonstrated that nicotinamide promoted pancreatic progenitor differentiation in chemically defined conditions, but it did not act through either niacin-associated metabolism or the inhibition of PARP and SIRT pathways. In contrast, nicotinamide modulated differentiation through CK1 and ROCK inhibition. We demonstrated that CK1 inhibitors promoted the generation of PDX1/NKX6.1 double-positive pancreatic progenitor cells. shRNA knockdown revealed that the inhibition of CK1α and CK1ε promoted pancreatic progenitor differentiation. We then showed that nicotinamide also improved pancreatic progenitor differentiation through ROCK inhibition. Finally, RNA-seq data showed that CK1 and ROCK inhibition led to pancreatic gene expression, similar to nicotinamide treatment. Conclusions In this report, we revealed that nicotinamide promotes generation of pancreatic progenitors from hESCs through CK1 and ROCK inhibition. Furthermore, we discovered the novel role of CK1 in pancreatic cell fate determination.

scholarly journals POGZ controls embryonic stem cell self-renewal and pluripotency by association with esBAF and HP1

2021 ◽  
Author(s):  
Xiaoyun Sun ◽  
Linxi Cheng ◽  
Yuhua Sun
Keyword(s):  
Cell Fate ◽  
Neurodevelopmental Disorders ◽  
Embryonic Stem ◽  
Autism Spectrum ◽  
Specific Gene ◽  
Open Chromatin ◽  
Cell Fate Determination ◽  
Chromatin Remodeler ◽  
Self Renewal ◽  
Fate Determination

AbstractPOGZ, which encodes a multi-domain transcription factor, has been found frequently mutated in neurodevelopmental disorders, particularly autism spectrum disorder (ASD) and intellectual disability (ID). However, little is known about its function in ESC self-renewal and pluripotency, cell fate determination as well as in transcriptional regulation. Here, using embryonic stem cells (ESCs) as model, we show that POGZ plays key roles in the maintenance of ESC and cell fate determination by association with the SWI-SNW chromatin remodeler complex and heterochromatin protein 1 (HP1) proteins. POGZ is essential for the maintenance of ESC undifferentiated state, and loss of POGZ leads to ESC differentiation, likely by up-regulation of primitive endoderm and mesoderm lineage genes and by down-regulation of pluripotency-related genes. Mechanistically, POGZ may control ESC-specific gene expression by association with chromatin remodeler complex esBAF and HP1s, and they can form a PBH triplex. POGZ functions primarily to maintain an open chromatin, as loss of POGZ leads to a reduced chromatin accessibility. Regulation of chromatin under control of POGZ depends on esBAF complex. POGZ is extensively co-localized with OCT4/NANOG genome wide. Taken together, we propose that POGZ is a pluripotency-associated factor, and its absence in ESCs causes failure to maintain a proper ESC-specific chromatin state and transcriptional circuitry of pluripotency, which eventually leads to ESC self-renewal and pluripotency defects. Our work provides important insights into the role of POGZ in ESC self-renewal and pluripotency as well as regulation of transcription, which will be useful for understanding the etiology of neurodevelopmental disorders by POGZ mutation.

scholarly journals Regulated Fluctuations in Nanog Expression Mediate Cell Fate Decisions in Embryonic Stem Cells

PLoS Biology ◽  
2009 ◽  
Vol 7 (7) ◽  
pp. e1000149 ◽  
Author(s):  
Tibor Kalmar ◽  
Chea Lim ◽  
Penelope Hayward ◽  
Silvia Muñoz-Descalzo ◽  
Jennifer Nichols ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Cell Fate ◽  
Embryonic Stem ◽  
Cell Fate Decisions ◽  
Nanog Expression ◽  
Mediate Cell

Effect of NANOG overexpression on porcine embryonic development and pluripotent embryonic stem cell formation in vitro

Zygote ◽  
2021 ◽  
pp. 1-6
Author(s):  
Gerelchimeg Bou ◽  
Shimeng Guo ◽  
Jia Guo ◽  
Zhuang Chai ◽  
Jianchao Zhao ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Embryonic Stem Cells ◽  
Embryonic Stem Cell ◽  
Embryonic Stem Cell Line ◽  
Embryonic Stem ◽  
Cell Clones ◽  
Morula Stage ◽  
Pluripotent Embryonic Stem Cell

Summary The efficiency of establishing pig pluripotent embryonic stem cell clones from blastocysts is still low. The transcription factor Nanog plays an important role in maintaining the pluripotency of mouse and human embryonic stem cells. Adequate activation of Nanog has been reported to increase the efficiency of establishing mouse embryonic stem cells from 3.5 day embryos. In mouse, Nanog starts to be strongly expressed as early as the morula stage, whereas in porcine NANOG starts to be strongly expressed by the late blastocyst stage. Therefore, here we investigated both the effect of expressing NANOG on porcine embryos early from the morula stage and the efficiency of porcine pluripotent embryonic stem cell clone formation. Compared with intact porcine embryos, NANOG overexpression induced a lower blastocyst rate, and did not show any advantages for embryo development and pluripotent embryonic stem cell line formation. These results indicated that, although NANOG is important pluripotent factor, NANOG overexpression is unnecessary for the initial formation of porcine pluripotent embryonic stem cell clones in vitro.

scholarly journals LncRNA Platr22 promotes super-enhancer activity and stem cell pluripotency

2020 ◽  
Author(s):  
Pixi Yan ◽  
J Yuyang Lu ◽  
Jing Niu ◽  
Juntao Gao ◽  
Michael Q Zhang ◽  
...  
Keyword(s):  
Cell Fate ◽  
Noncoding Rna ◽  
Embryonic Stem ◽  
Cell Fate Determination ◽  
Enhancer Activity ◽  
Lncrna Gene ◽  
Fate Determination ◽  
Super Enhancer ◽  
Stem Cell Pluripotency ◽  
Active Transcription

Abstract Super-enhancers (SEs) comprise large clusters of enhancers, which are co-occupied by multiple lineage-specific and master transcription factors, and play pivotal roles in regulating gene expression and cell fate determination. However, it is still largely unknown whether and how SEs are regulated by the non-coding portion of the genome. Here, through genome-wide analysis, we found that long noncoding RNA (lncRNA) genes preferentially lie next to SEs. In mouse embryonic stem cells (mESCs), depletion of SE-associated lncRNA transcripts dysregulated the activity of their nearby SEs. Specifically, we revealed a critical regulatory role of the lncRNA gene Platr22 in modulating the activity of a nearby SE and the expression of the nearby pluripotency regulator ZFP281. Through these regulatory events, Platr22 contributes to pluripotency maintenance and proper differentiation of mESCs. Mechanistically, Platr22 transcripts coat chromatin near the SE region and interact with DDX5 and hnRNP-L. DDX5 further recruits p300 and other factors related to active transcription. We propose that these factors assemble into a transcription hub, thus promoting an open and active epigenetic chromatin state. Our study highlights an unanticipated role for a class of lncRNAs in epigenetically controlling the activity and vulnerability to perturbation of nearby SEs for cell fate determination.

scholarly journals The L-type Ca2+ Channels Blocker Nifedipine Represses Mesodermal Fate Determination in Murine Embryonic Stem Cells

PLoS ONE ◽  
2013 ◽  
Vol 8 (1) ◽  
pp. e53407 ◽  
Author(s):  
Filomain Nguemo ◽  
Bernd K. Fleischmann ◽  
Manoj K. Gupta ◽  
Tomo Šarić ◽  
Daniela Malan ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Fate Determination ◽  
Murine Embryonic Stem Cells ◽  
Ca2 Channels

289 DERIVATION OF MOUSE EMBRYONIC STEM CELL FROM C57BL/6/EGFP STRAIN WITH FETAL CALF SERUM AND KNOCKOUT SERUM REPLACEMENT® SUPPLEMENTATION MEDIUM

2011 ◽  
Vol 23 (1) ◽  
pp. 242
Author(s):  
B. C. S. Campanha ◽  
C. S. Oliveira ◽  
D. M. Souza ◽  
C. P. Godoi ◽  
H. Fernandes ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Calf Serum ◽  
Embryonic Stem ◽  
Mouse Embryonic Stem Cell ◽  
Serum Replacement ◽  
Tetraploid Complementation ◽  
Murine Fibroblast ◽  
Esc Derivation ◽  
Knockout Serum Replacement

Embryonic stem cells (ESC) have been used in attempts to obtain specific tissues or even individuals. Embryonic stem cells are pluripotent, allowing the differentiation of cell types from 3 germ layers. The establishment of a stable lineage of ESC is a valuable tool; however, some strains of mice are less permissive to ESC derivation or generation of chimeric animals (e.g. C57BL/6). Supplementation of culture medium with FCS, in the ESC derivation, may influence the potentiality to derivation or use of these strains in tetraploid complementation assays (Sato et al. 2009 Tsukuba Res. Inst. 47, 414–422). Thus, its replacement was carried out using knockout serum replacement (KSR®) to minimize the deleterious action of serum (Wang et al. 2007 Inst. of Biotech. 23, 269–272). Embryos were obtained from 5 females of lineage C57BL6/EGFP, aged between 21 and 30 days and weighing ∼35 g, and superstimulated according (Mancini et al. 2008 Transg. Res. 17, 1015). The animals were placed for mating with fertile males of the same strain in a proportion of one to one (male:female). The copulation was confirmed by plug vaginal (0.5 days postcopulation). Embryo recovery was performed 3.5 to 4.0 days postcopulation to obtain expanded (EB) or hatched blastocysts (HB). Zona pellucida was removed from EB with the aid of pronase solution, and the whole embryos (n = 8) were placed on a 4-well dish pretreated with pig skin gelatin 0.1%, under murine fibroblast primary in DMEM medium supplemented with 7.5% FCS and 7.5% KSR®, 10 mM βmercaptoetanol, 1 mM sodium pyruvate, 2 mM L-glutamine, and 83.4 mg mL–1 amikacin for 24 h. After this period, the medium was replaced by DMEM supplemented with 15% KSR®. The colonies began to grow between 3 and 6 days after in vitro culture of the embryos. Once established, the colony was picked and placed into new plates containing murine fibroblast primary every 48 to 72 h. After 14 days, the derivation was confirmed with some proved pluripotency markers by immunofluorescence (Oct3/4, SSEA-1, and Nanog) and karyotyping for ploidy detection. The reaction was positive for all tested markers in addition to the detection of the endogenous fluorescence from EGFP protein itself (C57BL/6EGFP origin). It was concluded that ESC derivation with partial serum replacement and using a less permissive strain such as C57BL/6EGFP is feasible, although with a reduced success rate (12.5%; i.e. 1 lineage – named BCM04 – from 8 attempts). Fellowships and grants were received from FAPESP, Brazil: 09/15919-4 (BCSC), 09/16254-6 (DMS), 09/17605-7 (CPG), 06/06491-2 (MFGN), and 07/07705-9 (MFGN).

scholarly journals SIRT1 induces the adipogenic differentiation of mouse embryonic stem cells by regulating RA-induced RAR expression via NCOR1 acetylation

2020 ◽  
Vol 44 ◽  
pp. 101771 ◽  
Author(s):  
Yu Jin Jung ◽  
Woong Park ◽  
Jeong Mi Noh ◽  
Kyung Pyo Kang ◽  
Tung Nguyen-Thanh ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Adipogenic Differentiation ◽  
Embryonic Stem ◽  
Mouse Embryonic Stem Cells
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