Identification of differentially expressed genes in mouse embryonic stem cell under hypoxia
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Genes & Genomics https://doi.org/10.1007/s13258-020-01009-4
RESEARCH ARTICLE
Identification of differentially expressed genes in mouse embryonic stem cell under hypoxia Su Jung Hwang1,2 · Hyo‑Jong Lee1,2 Received: 22 June 2020 / Accepted: 7 October 2020 © The Genetics Society of Korea 2020
Abstract Background Under hypoxia, mouse embryonic stem cells (mESCs) lose the ability to self-renew and begin to differentiate through down-regulation of LIFR-STAT3 pathway via hypoxia-inducible factor-1α (HIF-1α). However, it remains largely unknown what kinds of factors are involved in hypoxia-induced differentiation of mESCs. Purpose This study aims to identify the differentially expressed genes (DEGs) in early differentiation of mESCs under hypoxia. Methods Here we utilized a Genefishing t echniqueTM to discover the new DEGs during hypoxia-induced early differentiation in CCE mESCs. Next, we investigated the role of DEGs using morphological observation, alkaline phosphatase (ALP) assay, STAT3 activation analysis, and biomarkers analysis for stemness. Results We detected 19 DEGs under hypoxia and performed cloning with sequencing in six genes. We confirmed the expression patterns of five DEGs including H2afz and GOT1 by realtime PCR assay. Among them, H2afz was significantly decreased under hypoxia, depending on HIF-1α. H2afz-overexpressing CCE mESCs maintained their ALP activity and stem cell markers (Nanog and Rex1), even in hypoxic condition. On the other hand, the early differentiation markers such as FGF5 and STAT5a, which had been increased in hypoxic conditions, were reduced by H2afz overexpression. Conclusion We discovered that H2afz could be a new target gene that functions in hypoxia-induced differentiation in mESCs and have revealed that it is involved in maintaining the pluripotency of mESCs in the early stages of differentiation. These findings will provide insights into mechanisms of hypoxia-mediated differentiation of mESCs during early development. Keywords Embryonic stem cell · DEGs · Hypoxia · HIF-1α · H2afz
Introduction Embryonic stem cells (mESCs) have been established previously from the inner cell mass of mouse blastocysts and have a potential to differentiate into any type of cell, including endothelial cells (Li et al. 2007), adipocytes (Bost et al. 2002), neurons (Schrenk-Siemens et al. 2008), and smooth muscle cells (Du et al. 2004; Yu and Thomson 2008). For these facts, mESCs have been widely used to study the * Hyo‑Jong Lee [email protected] 1
College of Pharmacy and Inje Institute of Pharmaceutical Sciences and Research, Inje University, 197 Inje‑ro, Gimhae, Gyungnam 50834, South Korea
School of Pharmacy, Sungkyunkwan University, 2066 Seobu‑ro, Jangan‑gu, Suwon, Gyeonggi‑do 16419, South Korea
2
early mammalian organogenesis for the development of cell therapy (Evans and Kaufman 1981). When mESCs were cultured with leukemia inhibitory factor (LIF), mESCs can be maintained indefinitely in a self-renewing state through LIF receptor (LIFR) and the signal transducer and activator o
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