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解析扩增人羊水干细胞亚群的异质性。

Unravelling Heterogeneity of Amplified Human Amniotic Fluid Stem Cells Sub-Populations.

机构信息

Department of Biomedical, Metabolic and Neural Sciences, University of Modena and Reggio Emilia, 41124 Modena, Italy.

Cellular Signalling Laboratory, Department of Biomedical and Neuromotor Sciences, University of Bologna, 40125 Bologna, Italy.

出版信息

Cells. 2021 Jan 15;10(1):158. doi: 10.3390/cells10010158.

DOI:10.3390/cells10010158
PMID:33467440
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7830644/
Abstract

Human amniotic fluid stem cells (hAFSCs) are broadly multipotent immature progenitor cells with high self-renewal and no tumorigenic properties. These cells, even amplified, present very variable morphology, density, intracellular composition and stemness potential, and this heterogeneity can hinder their characterization and potential use in regenerative medicine. Celector (Stem Sel ltd.) is a new technology that exploits the Non-Equilibrium Earth Gravity Assisted Field Flow Fractionation principles to characterize and label-free sort stem cells based on their solely physical characteristics without any manipulation. Viable cells are collected and used for further studies or direct applications. In order to understand the intrapopulation heterogeneity, various fractions of hAFSCs were isolated using the Celector profile and live imaging feature. The gene expression profile of each fraction was analysed using whole-transcriptome sequencing (RNAseq). Gene Set Enrichment Analysis identified significant differential expression in pathways related to Stemness, DNA repair, E2F targets, G2M checkpoint, hypoxia, EM transition, mTORC1 signalling, Unfold Protein Response and p53 signalling. These differences were validated by RT-PCR, immunofluorescence and differentiation assays. Interestingly, the different fractions showed distinct and unique stemness properties. These results suggest the existence of deep intra-population differences that can influence the stemness profile of hAFSCs. This study represents a proof-of-concept of the importance of selecting certain cellular fractions with the highest potential to use in regenerative medicine.

摘要

人羊水干细胞（hAFSCs）是一种广泛的多能未成熟祖细胞，具有高自我更新能力和无致瘤性。这些细胞即使扩增，也表现出非常不同的形态、密度、细胞内成分和干细胞特性，这种异质性可能会阻碍其表征和在再生医学中的潜在应用。

Celector（Stem Sel ltd.）是一种新技术，利用非平衡地球重力辅助场流分离原理，根据细胞的物理特性对干细胞进行无标记特征和分选，而无需任何操作。收集有活力的细胞用于进一步的研究或直接应用。为了了解细胞群体内的异质性，使用 Celector 图谱和活细胞成像功能对 hAFSCs 的各个部分进行分离。使用全转录组测序（RNAseq）分析每个部分的基因表达谱。基因集富集分析确定了与干细胞特性、DNA 修复、E2F 靶标、G2M 检查点、缺氧、EM 转化、mTORC1 信号、未折叠蛋白反应和 p53 信号相关的途径中的显著差异表达。通过 RT-PCR、免疫荧光和分化实验验证了这些差异。

有趣的是，不同的部分显示出独特的干细胞特性。这些结果表明存在深刻的细胞群体内差异，这可能会影响 hAFSCs 的干细胞特性。这项研究代表了选择具有最高再生医学应用潜力的特定细胞部分的重要性的概念验证。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/de6f/7830644/b85010046019/cells-10-00158-g007.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/de6f/7830644/f6d786c9f540/cells-10-00158-g001.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/de6f/7830644/b85010046019/cells-10-00158-g007.jpg

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