Single-cell RNA-seq highlights heterogeneity in human primary Wharton's jelly mesenchymal stem/stromal cells cultured in vitro
Single-cell RNA-seq highlights heterogeneity in human primary Wharton's jelly mesenchymal stem/stromal cells cultured in vitro
复制标题
单细胞RNA-seq突出了体外培养的人原代沃顿胶质间充质干细胞/基质细胞的异质性
DOI：
10.1186/s13287-020-01660-4
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发表时间：
2020-04-06
影响因子：
7.5
通讯作者：
Zhang, Xi
中科院分区：
文献类型：
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作者：
Sun, Changbin;Wang, Lei;Zhang, Xi
背景间充质干/基质细胞（Mesenchymal stem/stromal cells，MSCs）是一种多能干细胞，在再生医学和免疫调节方面具有广阔的应用前景。然而，体外培养的MSC表现出功能异质性。定义MSC异质性的潜在分子机制仍不清楚。方法采用单细胞RNA测序技术（scRNA-seq）对3名供体体外培养的人原代Wharton's骨髓间充质干细胞（WJMSCs）进行基因表达谱分析。我们还基于scRNA-seq数据分离了CD 142（+）和CD 142（-）WJMSCs，并比较了它们的体外增殖能力和“伤口愈合”潜力。同时，我们分析了公开可用的脂肪来源的MSC（ADMSCs）scRNA-seq数据，并在单细胞水平上进行了WJMSC和ADMSCs之间的转录组比较。结果从WJMSCs中获得的高度可变基因（HVGs）的GO富集分析显示，这些基因在细胞外区域显著富集，具有结合功能，参与发育过程、信号转导、细胞增殖等。通路分析显示，这些HVGs与经典MSCs的功能特征相关，如趋化因子和细胞因子信号转导介导的炎症、整合素信号转导、和血管生成。在回归出批次和细胞周期的影响后，这些HVG用于降维和聚类分析，以确定候选亚群。差异表达基因分析显示存在几种不同的MSC亚群，其表现出与增殖、发育和炎症反应相关的不同功能特征。与我们的数据一致，分选的CD 142（+）和CD 142（-）WJMSCs显示出不同的增殖能力以及“伤口愈合”潜力。尽管WJMSC和ADMSC来源于不同组织，表现出不同的分化能力，但它们的HVG在很大程度上重叠，并且具有相似的功能富集。结论在MSC中鉴定的HVG与MSC的经典功能相关。关于治疗潜力，这些基因与MSC临床应用理论上所基于的功能特征相关，例如发育和炎症反应。总之，这些HVG具有用作进一步效价相关性研究的候选标记物的潜力。
Background Mesenchymal stem/stromal cells (MSCs) are multipotent cells with a promising application potential in regenerative medicine and immunomodulation. However, MSCs cultured in vitro exhibit functional heterogeneity. The underlying molecular mechanisms that define MSC heterogeneity remain unclear. Methods We investigated the gene expression profile via single-cell RNA sequencing (scRNA-seq) of human primary Wharton's jelly-derived MSCs (WJMSCs) cultured in vitro from three donors. We also isolated CD142(+) and CD142(-) WJMSCs based on scRNA-seq data and compared their proliferation capacity and "wound healing" potential in vitro. Meanwhile, we analyzed publicly available adipose-derived MSC (ADMSCs) scRNA-seq data and performed transcriptome comparison between WJMSCs and ADMSCs at the single-cell level. Results GO enrichment analysis of highly variable genes (HVGs) obtained from WJMSCs revealed that these genes are significantly enriched in extracellular region with binding function, involved in developmental process, signal transduction, cell proliferation, etc. Pathway analysis showed that these HVGs are associated with functional characteristics of classic MSCs, such as inflammation mediated by chemokine and cytokine signaling, integrin signaling, and angiogenesis. After regressing out the batch and cell cycle effects, these HVGs were used for dimension reduction and clustering analysis to identify candidate subpopulations. Differentially expressed gene analysis revealed the existence of several distinct subpopulations of MSCs that exhibit diverse functional characteristics related to proliferation, development, and inflammation response. In line with our data, sorted CD142(+) and CD142(-) WJMSCs showed distinct proliferation capacity as well as "wound healing" potential. Although WJMSCs and ADMSCs were derived from different tissues and were displaying different differentiation potencies, their HVGs were largely overlapped and had similar functional enrichment. Conclusion HVGs identified in MSCs are associated with classic MSC function. Regarding therapeutic potential, these genes are associated with functional characteristics, on which the MSC clinical application were theoretically based, such as development and inflammation response. Altogether, these HVGs hold the potential to be used as candidate markers for further potency association studies.