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微小 RNA 在人类诱导多能干细胞 (hiPSC) 早期软骨生成中的作用。

The Role of MicroRNAs in Early Chondrogenesis of Human Induced Pluripotent Stem Cells (hiPSCs).

机构信息

Radiobiology Lab, Greater Poland Cancer Centre, Garbary 15th Street, 61-866 Poznan, Poland.

Department of Histology and Embryology, Poznan University of Medical Sciences, Swiecickiego 6 Street, 60-781 Poznan, Poland.

出版信息

Int J Mol Sci. 2019 Sep 5;20(18):4371. doi: 10.3390/ijms20184371.

Abstract

Human induced pluripotent stem cells (hiPSCs) play an important role in research regarding regenerative medicine. Particularly, chondrocytes differentiated from hiPSCs seems to be a promising solution for patients suffering from osteoarthritis. We decided to perform chondrogenesis in a three-week monolayer culture. Based on transcriptome analysis, hiPSC-derived chondrocytes (ChiPS) demonstrate the gene expression profile of cells from early chondrogenesis. Chondrogenic progenitors obtained by our group are characterized by significantly high expression of Hox genes, strongly upregulated during limb formation and morphogenesis. There are scanty literature data concerning the role of microRNAs in early chondrogenesis, especially in chondrogenic differentiation of hiPSCs. The main aim of this study was to investigate the microRNA expression profile and to select microRNAs (miRNAs) taking part in early chondrogenesis. Our findings allowed for selection crucial miRNAs engaged in both diminishing pluripotency state and chondrogenic process (inter alia hsa-miR-525-5p, hsa-miR-520c-3p, hsa-miR-628-3p, hsa-miR-196b-star, hsa-miR-629-star, hsa-miR-517b, has-miR-187). These miRNAs regulate early chondrogenic genes such as: , , , . These results were confirmed by RT-qPCR analysis. This work contributes to a better understanding of the role of miRNAs directly involved in chondrogenic differentiation of hiPSCs. These data may result in the establishment of a more efficient protocol of obtaining chondrocyte-like cells from hiPSCs.

摘要

人诱导多能干细胞(hiPSCs)在再生医学研究中发挥着重要作用。特别是,hiPSC 分化而来的软骨细胞似乎是治疗骨关节炎患者的有前途的方法。我们决定在为期三周的单层培养中进行软骨生成。基于转录组分析,hiPSC 来源的软骨细胞(ChiPS)表现出早期软骨生成细胞的基因表达谱。我们小组获得的软骨祖细胞的特征是 Hox 基因的表达水平显著升高,在肢体形成和形态发生过程中强烈上调。关于 microRNAs 在早期软骨生成中的作用,特别是在 hiPSC 的软骨分化中的作用,文献数据很少。本研究的主要目的是研究 microRNA 的表达谱,并选择参与早期软骨生成的 microRNAs。我们的研究结果允许选择参与减少多能性状态和软骨生成过程的关键 microRNAs(例如 hsa-miR-525-5p、hsa-miR-520c-3p、hsa-miR-628-3p、hsa-miR-196b-star、hsa-miR-629-star、hsa-miR-517b、has-miR-187)。这些 microRNAs 调节早期软骨生成基因,如: 、 、 、 。这些结果通过 RT-qPCR 分析得到了证实。这项工作有助于更好地理解直接参与 hiPSC 软骨分化的 microRNAs 的作用。这些数据可能导致建立一种从 hiPSC 获得类软骨细胞的更有效的方案。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d30a/6770352/0731a03b5bde/ijms-20-04371-g001.jpg

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