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诱导多能干细胞(iPSC)衍生的胰腺β样细胞分化的转录组分析。

Transcriptome Analysis of Induced Pluripotent Stem Cell (iPSC)-derived Pancreatic β-like Cell Differentiation.

机构信息

1 Department of General Surgery, Affiliated Hospital of Nantong University, Nantong, Jiangsu Province, China.

2 Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, Nantong, Jiangsu Province, China.

出版信息

Cell Transplant. 2017 Aug;26(8):1380-1391. doi: 10.1177/0963689717720281.

Abstract

Diabetes affects millions of people worldwide, and β-cell replacement is one of the promising new strategies for treatment. Induced pluripotent stem cells (iPSCs) can differentiate into any cell type, including pancreatic β cells, providing a potential treatment for diabetes. However, the molecular mechanisms underlying the differentiation of iPSC-derived β cells have not yet been fully elucidated. Here, we generated pancreatic β-like cells from mouse iPSCs using a 3-step protocol and performed deep RNA sequencing to get a transcriptional landscape of iPSC-derived pancreatic β-like cells during the selective differentiation period. We then focused on the differentially expressed genes (DEGs) during the time course of the differentiation period, and these genes underwent Gene Ontology annotation and Kyoto Encyclopedia of Genes and Genomes pathway analysis. In addition, gene-act networks were constructed for these DEGs, and the expression of pivotal genes detected by quantitative real-time polymerase chain reaction was well correlated with RNA sequence (RNA-seq). Overall, our study provides valuable information regarding the transcriptome changes in β cells derived from iPSCs during differentiation, elucidates the biological process and pathways underlying β-cell differentiation, and promotes the identification and functional analysis of potential genes that could be used for improving functional β-cell generation from iPSCs.

摘要

糖尿病影响着全球数百万人,β 细胞替代是治疗的一种有前途的新策略。诱导多能干细胞(iPSCs)可以分化为任何细胞类型,包括胰腺β细胞,为糖尿病的治疗提供了一种潜在的方法。然而,iPSC 来源的β 细胞分化的分子机制尚未完全阐明。在这里,我们使用 3 步方案从鼠 iPSCs 中产生胰腺β样细胞,并进行深度 RNA 测序,以获得选择性分化期间 iPSC 来源的胰腺β样细胞的转录组图谱。然后,我们专注于分化期间时间过程中的差异表达基因(DEGs),这些基因进行了基因本体论注释和京都基因与基因组百科全书通路分析。此外,构建了这些 DEGs 的基因活性网络,通过定量实时聚合酶链反应检测的关键基因的表达与 RNA 序列(RNA-seq)很好地相关。总的来说,我们的研究提供了关于 iPSC 来源的β 细胞在分化过程中转录组变化的有价值的信息,阐明了β 细胞分化的生物学过程和途径,并促进了对潜在基因的鉴定和功能分析,这些基因可能用于改善 iPSC 来源的功能性β 细胞的生成。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b7dc/5680972/a9ffb0391088/10.1177_0963689717720281-fig1.jpg

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