Single cell studies of mouse embryonic stem cell (mESC) differentiation by electrical impedance measurements in a microfluidic device.
Single cell studies of mouse embryonic stem cell (mESC) differentiation by electrical impedance measurements in a microfluidic device.
复制标题
DOI：
10.1016/j.bios.2016.02.069
复制
发表时间：
2016-07-15
影响因子：
12.6
通讯作者：
Seshia AA
中科院分区：
文献类型：
--
作者：
Zhou Y;Basu S;Laue E;Seshia AA
细胞的生物群体显示出相当大的细胞间变异性。单细胞的研究和细胞异质性的分析被认为是理解生物过程，如干细胞分化和癌症发展的关键。芯片实验室技术的最新进展已经允许在微流体通道中捕获单细胞，具有精确的环境控制和高通量实验的可能性，同时使用最少的样品和试剂。近年来，电阻抗谱等无标记技术已成为研究细胞特性的非侵入性方法。在这项研究中，我们设计并制造了一种微流体装置，该装置将单细胞在预定义位置的流体动力学捕获与在同一装置内运行电阻抗测量的能力相结合。对小鼠胚胎干细胞（mESCs）在分化过程中不同状态（t= 0h、24h和48h）的电参数进行了测量，并定量分析了细胞在分化过程中电参数的变化。在细胞分化过程中发现细胞阻抗的幅度显著增加，这可以归因于细胞大小的增加。对测量结果的分析表明，在此过程中，核质比降低。与未分化（0 h）和完全分化（48 h）状态的细胞相比，当细胞处于过渡状态（24 h）时，观察到细胞异质性的程度最高。该设备能够高效地捕获单个细胞，并提供单个细胞的灵敏、无标记电阻抗测量，从而能够定量分析其物理状态以及研究细胞群体的相关异质性。微流体装置结合了流体动力学捕获和阻抗谱。在分化周期中捕获和监测小鼠胚胎干细胞。在分化过程中观察到细胞电参数的显着变化。定量分析了分化过程中细胞异质性的程度。高灵敏度的无标记测量的电细胞标志物证明。
Biological populations of cells show considerable cell-to-cell variability. Study of single cells and analysis of cell heterogeneity are considered to be critical in understanding biological processes such as stem cell differentiation and cancer development. Recent advances in lab-on-a-chip techniques have allowed single-cell capture in microfluidic channels with the possibility of precise environmental control and high throughput of experiments with minimal usage of samples and reagents. In recent years, label-free techniques such as electrical impedance spectroscopy have emerged as a non-invasive approach to studying cell properties. In this study, we have designed and fabricated a microfluidic device that combines hydrodynamic trapping of single cells in pre-defined locations with the capability of running electrical impedance measurements within the same device. We have measured mouse embryonic stem cells (mESCs) at different states during differentiation (t=0 h, 24 h and 48 h) and quantitatively analysed the changes in electrical parameters of cells during differentiation. A marked increase in the magnitude of the cell impedance is found during cell differentiation, which can be attributed to an increase in cell size. The analysis of the measurements shows that the nucleus-to-cytoplasm ratio decreases during this process. The degree of cell heterogeneity is observed to be the highest when the cells are at the transition state (24 h), compare with cells at undifferentiated (0 h) and fully differentiated (48 h) states. The device enables highly efficient single cell trapping and provides sensitive, label-free electrical impedance measurements of individual cells, enabling the possibility of quantitatively analysing their physical state as well as studying the associated heterogeneity of a cell population. Microfluidic device combines hydrodynamic trapping and impedance spectroscopy. Mouse embryonic stem cells trapped and monitored during the differentiation cycle. A marked variation in cell electrical parameters is observed during differentiation. The degree of cell heterogeneity during differentiation is quantitatively analysed. Highly sensitive label-free measurements of electrical cell markers demonstrated.