人孤雌胚胎干细胞向功能性肝细胞样细胞的分化。

IF 1.6 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY
Organogenesis Pub Date : 2020-10-01 Epub Date: 2020-11-25 DOI:10.1080/15476278.2020.1848237
Rui Liang, Zhiqiang Wang, Xiangyang Kong, Xiaoxiao Xiao, Tianxing Chen, Hui Yang, Ying Li, Xingqi Zhao
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引用次数: 2

摘要

干细胞和组织工程治疗急性肝衰竭(ALF)一直受到缺乏最佳细胞来源的限制。我们的目的是确定人类单性生殖胚胎干细胞(hPESCs)在ALF治疗策略开发中的适用性。我们研究了在体外受精(IVF)过程中通过自然激活获得的具有高全基因组SNP纯合性的人单性生殖胚胎干细胞(hPESCs)在体外通过单层平面定向分化为功能性肝细胞样细胞的能力。用激活素a、FGF-4、BMP-2、HGF、OSM、DEX和B27等诱导物在单层平板上诱导hPESCs 21 d。倒置显微镜下观察诱导21 d后的多角形细胞形态和双核细胞。RT-qPCR结果显示,诱导后肝细胞特异性基因如AFP、ALB、HNF4a、CYP3A4、SLCO1B3、ABCC2水平显著升高。免疫细胞化学检测显示CK18和Hepa在诱导细胞中表达。吲哚菁绿(ICG)染色表明细胞具有吸收和代谢染料的能力。细胞培养上清中标记蛋白和尿素的检测表明,诱导21 d后获得的细胞具有合成和分泌功能。诱导21 d后用透射电镜观察肝细胞的典型超微结构。结果表明,自然激活的hPESCs可通过单层平面诱导向肝细胞分化。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Differentiation of Human Parthenogenetic Embryonic Stem Cells into Functional Hepatocyte-like Cells.

Stem cell and tissue engineering-based therapies for acute liver failure (ALF) have been limited by the lack of an optimal cell source. We aimed to determine the suitability of human parthenogenetic embryonic stem cells (hPESCs) for the development of strategies to treat ALF. We studied the ability of human parthenogenetic embryonic stem cells (hPESCs) with high whole-genome SNP homozygosity, which were obtained by natural activation during in vitro fertilization (IVF), to differentiate into functional hepatocyte-like cells in vitro by monolayer plane orientation. hPESCs were induced on a single-layer flat plate for 21 d in complete medium with the inducers activin A, FGF-4, BMP-2, HGF, OSM, DEX, and B27. Polygonal cell morphology and binuclear cells were observed after 21 d of induction by using an inverted microscope. RT-qPCR results showed that the levels of hepatocyte-specific genes such as AFP, ALB, HNF4a, CYP3A4, SLCO1B3, and ABCC2 significantly increased after induction. Immunocytochemical assay showed CK18 and Hepa expression in the induced cells. Indocyanine green (ICG) staining showed that the cells had the ability to absorb and metabolize dyes. Detection of marker proteins and urea in cell culture supernatants showed that the cells obtained after 21 d of induction had synthetic and secretory functions. The typical ultrastructure of liver cells was observed using TEM after 21 d of induction. The results indicate that naturally activated hPESCs can be induced to differentiate into hepatocellular cells by monolayer planar induction.

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来源期刊
Organogenesis
Organogenesis BIOCHEMISTRY & MOLECULAR BIOLOGY-DEVELOPMENTAL BIOLOGY
CiteScore
4.10
自引率
4.30%
发文量
6
审稿时长
>12 weeks
期刊介绍: Organogenesis is a peer-reviewed journal, available in print and online, that publishes significant advances on all aspects of organ development. The journal covers organogenesis in all multi-cellular organisms and also includes research into tissue engineering, artificial organs and organ substitutes. The overriding criteria for publication in Organogenesis are originality, scientific merit and general interest. The audience of the journal consists primarily of researchers and advanced students of anatomy, developmental biology and tissue engineering. The emphasis of the journal is on experimental papers (full-length and brief communications), but it will also publish reviews, hypotheses and commentaries. The Editors encourage the submission of addenda, which are essentially auto-commentaries on significant research recently published elsewhere with additional insights, new interpretations or speculations on a relevant topic. If you have interesting data or an original hypothesis about organ development or artificial organs, please send a pre-submission inquiry to the Editor-in-Chief. You will normally receive a reply within days. All manuscripts will be subjected to peer review, and accepted manuscripts will be posted to the electronic site of the journal immediately and will appear in print at the earliest opportunity thereafter.
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