Covariation of Pluripotency Markers and Biomechanical Properties in Mouse Embryonic Stem Cells.
Covariation of Pluripotency Markers and Biomechanical Properties in Mouse Embryonic Stem Cells.
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DOI：
10.3389/fcell.2022.858884
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发表时间：
2022
影响因子：
5.5
通讯作者：
Lee, David A.
中科院分区：
文献类型：
--
作者：
Brookes, Oliver;Thorpe, Stephen D.;Rigby Evans, Olga;Keeling, Michael C.;Lee, David A.
多能细胞作为研究模型、再生医学疗法和实验室培养肉等新应用的分化细胞材料的来源，受到了极大的关注。因此，更多地了解多能性状态并控制其分化是可取的。近年来，生物力学特性在指导细胞命运和细胞行为中的作用越来越得到充分的描述。然而，在体细胞中控制细胞形态和力学性质的许多机制在多能细胞中是缺失的。我们利用小鼠胚胎干细胞中生物力学特性和多能性基因表达的自然发生的变化来研究这些参数之间的关系。我们观察到REx1-GFP表达报告线的相当大的变异，并发现这种变异与细胞扩散形态没有明显的相关性，无论是在逐个参数的基础上，还是当使用从四个单独标准的主成分分析得出的组合度量时，这种变异与细胞扩散形态没有明显的相关性，这是由圆形、费雷特比率、相位对比亮度或细胞扩散面积决定的。我们进一步证实，细胞体积与REx1-GFP的表达没有协同变化。有趣的是，我们确实发现，容易被温和搅拌分离的细胞亚群共同表现出更高的Nanog表达和更低的LMNA表达，这表明多能性基因的表达增强可能与与底物的粘附性降低有关。此外，原子力显微镜和定量荧光成像显示细胞硬度和REx1-GFP报告表达之间存在联系。表达高水平REx1-GFP的细胞始终具有相对较低的刚性，而表达低水平REx1-GFP的细胞倾向于较高的硬度值。这些观察表明多能性基因表达和生物力学特性之间存在一定的相互作用，但也支持细胞培养机制和细胞生物力学特性之间的其他相互作用，这些相互作用独立于支持多能性的核心转录网络而发生。
Pluripotent cells are subject to much interest as a source of differentiated cellular material for research models, regenerative medical therapies and novel applications such as lab-cultured meat. Greater understanding of the pluripotent state and control over its differentiation is therefore desirable. The role of biomechanical properties in directing cell fate and cell behavior has been increasingly well described in recent years. However, many of the mechanisms which control cell morphology and mechanical properties in somatic cells are absent from pluripotent cells. We leveraged naturally occurring variation in biomechanical properties and expression of pluripotency genes in murine ESCs to investigate the relationship between these parameters. We observed considerable variation in a Rex1-GFP expression reporter line and found that this variation showed no apparent correlation to cell spreading morphology as determined by circularity, Feret ratio, phase contrast brightness or cell spread area, either on a parameter-by-parameter basis, or when evaluated using a combined metric derived by principal component analysis from the four individual criteria. We further confirmed that cell volume does not co-vary with Rex1-GFP expression. Interestingly, we did find that a subpopulation of cells that were readily detached by gentle agitation collectively exhibited higher expression of Nanog, and reduced LmnA expression, suggesting that elevated pluripotency gene expression may correlate with reduced adhesion to the substrate. Furthermore, atomic force microscopy and quantitative fluorescent imaging revealed a connection between cell stiffness and Rex1-GFP reporter expression. Cells expressing high levels of Rex1-GFP are consistently of a relatively low stiffness, while cells with low levels of Rex1-GFP tend toward higher stiffness values. These observations indicate some interaction between pluripotency gene expression and biomechanical properties, but also support a strong role for other interactions between the cell culture regime and cellular biomechanical properties, occurring independently of the core transcriptional network that supports pluripotency.
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影响因子：
3.4
作者：
Poh, Yeh-Chuin;Chowdhury, Farhan;Wang, Ning
通讯作者：
Wang, Ning
影响因子：
8.8
作者：
Maffioletti SM;Sarcar S;Henderson ABH;Mannhardt I;Pinton L;Moyle LA;Steele-Stallard H;Cappellari O;Wells KE;Ferrari G;Mitchell JS;Tyzack GE;Kotiadis VN;Khedr M;Ragazzi M;Wang W;Duchen MR;Patani R;Zammit PS;Wells DJ;Eschenhagen T;Tedesco FS
通讯作者：
Tedesco FS
影响因子：
3.5
作者：
Gavara N;Chadwick RS
通讯作者：
Chadwick RS
影响因子：
3.7
作者：
Chowdhury F;Li Y;Poh YC;Yokohama-Tamaki T;Wang N;Tanaka TS
通讯作者：
Tanaka TS
影响因子：
5.6
作者：
Becker, Klaus A.;Ghule, Prachi N.;Stein, Gary S.
通讯作者：
Stein, Gary S.