Enhanced Induction of Definitive Endoderm Differentiation of Mouse Embryonic Stem Cells in Simulated Microgravity

Artificially generated pancreatic β-cells from pluripotent stem cells are expected for cell replacement therapy for type 1 diabetes. Several strategies are adopted to direct pluripotent stem cells toward pancreatic differentiation. However, a standard differentiation method for clinical application has not been established. It is important to develop more effective and safer methods for generating pancreatic β-cells without toxic or mutagenic chemicals. In the present study, we screened several endogenous factors involved in organ development to identify the factor, which induced the efficiency of pancreatic differentiation and found that treatment with erythropoietin (EPO) facilitated the differentiation of mouse embryonic stem cells (ESCs) into definitive endoderm. At an early stage of differentiation, EPO treatment significantly increased Sox17 gene expression, as a marker of the definitive endoderm. Contrary to the canonical function of EPO, it did not affect the levels of phosphorylated JAK2 and STAT5, but stimulated the phosphorylation of ERK1/2 and Akt. The MEK inhibitor U0126 significantly inhibited EPO-induced Sox17 expression. The differentiation of ESCs into definitive endoderm is an important step for the differentiation into pancreatic and other endodermal lineages. This study suggests a possible role of EPO in embryonic endodermal development and a new agent for directing the differentiation into endodermal lineages like pancreatic β-cells.

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Serum and non-serum method-induced differentiation of mouse embryonic stem cells into definitive endoderm

Academic Journal of Second Military Medical University ◽

10.3724/sp.j.1008.2013.00124 ◽

2013 ◽

Vol 33 (2) ◽

pp. 124-129

Author(s):

Yang-fang LI ◽

Yong LAN ◽

Ya-zhuo ZHANG ◽

Xin WANG ◽

Yi-ping HU

Keyword(s):

Stem Cells ◽

Embryonic Stem Cells ◽

Embryonic Stem ◽

Mouse Embryonic Stem Cells ◽

Definitive Endoderm ◽

Induced Differentiation

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Definitive Endoderm Differentiation of Human Embryonic Stem Cells Combined with Selective Elimination of Undifferentiated Cells by Methionine Deprivation

Methods in Molecular Biology - Human Embryonic Stem Cell Protocols ◽

10.1007/7651_2015_224 ◽

2015 ◽

pp. 205-212 ◽

Cited By ~ 2

Author(s):

Tomonori Tsuyama ◽

Nobuaki Shiraki ◽

Shoen Kume

Keyword(s):

Stem Cells ◽

Embryonic Stem Cells ◽

Human Embryonic Stem Cells ◽

Embryonic Stem ◽

Definitive Endoderm ◽

Endoderm Differentiation ◽

Selective Elimination ◽

Undifferentiated Cells

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Arginine methylation of METTL14 promotes RNA N6-methyladenosine modification and endoderm differentiation of mouse embryonic stem cells

Nature Communications ◽

10.1038/s41467-021-24035-6 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Xiaona Liu ◽

Hailong Wang ◽

Xueya Zhao ◽

Qizhi Luo ◽

Qingwen Wang ◽

...

Keyword(s):

Stem Cells ◽

Embryonic Stem Cells ◽

Mutant Mouse ◽

Embryonic Stem ◽

Arginine Methylation ◽

Mouse Embryonic Stem Cells ◽

Protein Methylation ◽

Dynamic Regulation ◽

Endoderm Differentiation

AbstractRNA N6-methyladenosine (m6A), the most abundant internal modification of mRNAs, plays key roles in human development and health. Post-translational methylation of proteins is often critical for the dynamic regulation of enzymatic activity. However, the role of methylation of the core methyltransferase METTL3/METTL14 in m6A regulation remains elusive. We find by mass spectrometry that METTL14 arginine 255 (R255) is methylated (R255me). Global mRNA m6A levels are greatly decreased in METTL14 R255K mutant mouse embryonic stem cells (mESCs). We further find that R255me greatly enhances the interaction of METTL3/METTL14 with WTAP and promotes the binding of the complex to substrate RNA. We show that protein arginine N-methyltransferases 1 (PRMT1) interacts with and methylates METTL14 at R255, and consistent with this, loss of PRMT1 reduces mRNA m6A modification globally. Lastly, we find that loss of R255me preferentially affects endoderm differentiation in mESCs. Collectively, our findings show that arginine methylation of METTL14 stabilizes the binding of the m6A methyltransferase complex to its substrate RNA, thereby promoting global m6A modification and mESC endoderm differentiation. This work highlights the crosstalk between protein methylation and RNA methylation in gene expression.

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