In vitro metabolic zonation through oxygen gradient on a chip

doi:10.1038/s41598-019-49412-6

KMS > 免疫化学研究所 > 特聘教授组 > Nicola Elvassore组

	In vitro metabolic zonation through oxygen gradient on a chip
	Tonon, Federica 1; Giobbe, Giovanni Giuseppe 2,3; Zambon, Alessandro 3; Luni, Camilla4 ; Gagliano, Onelia 3,6; Floreani, Annarosa 5; Grassi, Gabriele 1; Elvassore, Nicola2,3,4,6
	2019-09-19
发表期刊	SCIENTIFIC REPORTS
ISSN	2045-2322
卷号	9
发表状态	已发表
DOI	10.1038/s41598-019-49412-6
摘要	Among the multiple metabolic signals involved in the establishment of the hepatic zonation, oxygen could play a key role. Indeed, depending on hepatocyte position in the hepatic lobule, gene expression and metabolism are differently affected by the oxygen gradient present across the lobule. The aim of this study is to understand whether an oxygen gradient, generated in vitro in our developed device, is sufficient to instruct a functional metabolic zonation during the differentiation of human embryonic stem cells (hESCs) from endoderm toward terminally differentiated hepatocytes, thus mimicking the in vivo situation. For this purpose, a microfluidic device was designed for the generation of a stable oxygen gradient. The oxygen gradient was applied to differentiating hESCs at the pre-hepatoblast stage. The definitive endoderm and hepatic endoderm cells were characterized by the expression of the transcription factor SOX-17 and alpha-fetoprotein (AFP). Immature and mature hepatocytes were characterized by hepatocyte nuclear factor 4-alpha (HNF-4 alpha) and albumin (ALB) expression and also analyzed for cytochrome P450 (CYP3A4) zonation and glycogen accumulation through PAS staining. Metabolic zonated genes expression was assessed through quantitative real time PCR. Application of the oxygen gradient during differentiation induced zonated glycogen storage, which was higher in the hepatocytes grown in high pO(2) compared to those grown in low pO(2). The mRNA levels of glutamine synthetase (GLUL), beta-catenin (CTNNB) and its direct target cyclin Dl (CCND1) showed significantly higher expression in the cells grown in low pO(2) compared to those grown in high pO(2). On the contrary, carbamoyl-phosphate synthetase 1 (CPS1), ALB, the proliferative marker ki67 (MKI67) and cyclin A (CCNA) resulted to be significantly higher expressed in cells cultured in high pO(2) compared to those cultured in low pO(2). These results indicate that the oxygen gradient generated in our device can instruct the establishment of a functional metabolic zonation in differentiating hESCs. The possibility to obtain differentiated hepatocytes in vitro may allow in the future to deepen our knowledge about the physiology/pathology of hepatocytes in relation to the oxygen content.
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收录类别	SCI ; SCIE
语种	英语
资助项目	Oak Foundation Award[W1095/OCAY-14-191]
WOS研究方向	Science & Technology - Other Topics
WOS类目	Multidisciplinary Sciences
WOS记录号	WOS:000486567800033
出版者	NATURE PUBLISHING GROUP
WOS关键词	LIVER ; DIFFERENTIATION ; CELLS ; HEPATOCYTES ; MODEL
原始文献类型	Article
引用统计
文献类型	期刊论文
条目标识符	https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/80226
专题	免疫化学研究所_特聘教授组_生物工程学实验室
通讯作者	Elvassore, Nicola
作者单位	1.Univ Trieste, Dept Life Sci, I-34127 Trieste, Italy 2.UCL GOS Inst Child Hlth, Stem Cell & Regenerat Med Sect, London WC1N 1EH, England 3.Univ Padua, Dept Ind Engn, I-35131 Padua, Italy 4.ShanghaiTech Univ, SIAIS, Shanghai 201210, Peoples R China 5.Univ Padua, Dept Surg Oncol & Gastroenterol, I-35121 Padua, Italy 6.Venetian Inst Mol Med, I-35129 Padua, Italy
通讯作者单位	上海科技大学
推荐引用方式 GB/T 7714	Tonon, Federica,Giobbe, Giovanni Giuseppe,Zambon, Alessandro,et al. In vitro metabolic zonation through oxygen gradient on a chip[J]. SCIENTIFIC REPORTS,2019,9.
APA	Tonon, Federica.,Giobbe, Giovanni Giuseppe.,Zambon, Alessandro.,Luni, Camilla.,Gagliano, Onelia.,...&Elvassore, Nicola.(2019).In vitro metabolic zonation through oxygen gradient on a chip.SCIENTIFIC REPORTS,9.
MLA	Tonon, Federica,et al."In vitro metabolic zonation through oxygen gradient on a chip".SCIENTIFIC REPORTS 9(2019).