Mechanical disruption of mammalian cells in a microfluidic system and its numerical analysis based on computational fluid dynamics.
Mechanical disruption of mammalian cells in a microfluidic system and its numerical analysis based on computational fluid dynamics.
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微流体系统中哺乳动物细胞的机械破坏及其基于计算流体动力学的数值分析
DOI：
10.1039/c2lc20918g
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发表时间：
2011
期刊：
影响因子：
6.1
通讯作者：
Zeng AP
中科院分区：
文献类型：
--
作者：
Wurm M;Zeng AP
哺乳动物细胞的裂解是不同的芯片实验室样品制备方法的重要组成部分，这些方法旨在释放、分离和随后分析DNA、蛋白质或代谢物。特别是对于哺乳动物细胞活体代谢的分室分析，这种方法必须比代谢更新率更快，它应该不影响天然代谢物的浓度，并且理想地应该保持细胞细胞器不受损害。到目前为止，还没有这样的方法可用。为了有效、快速地机械破碎细胞，我们开发了一套微流控系统，并建立了描述该系统效率的数学模型。中国仓鼠卵巢(CHO)细胞通过两个连续的微孔阵列进行高效破碎。同时单元压缩和剪切导致了≥90%的破碎率，当样品流量Q=120μL−1/喷嘴通道时，对应的平均流体速度为13.3m S−1，喷嘴缝隙内的平均雷诺数为22.6。我们讨论了在微喷口阵列上由蜂窝状碎片造成的通道堵塞以及由此引起的流动不稳定性问题。将实验结果与计算流体力学模拟的结果进行了比较，确定了已知尺寸分布的CHO细胞群破裂的临界能量耗散率为4.7×108W m−3。基于计算流体动力学数据计算细胞破裂的模型可以应用于其他微观几何模型，以预测有意破坏或不希望发生的细胞损伤。
The lysis of mammalian cells is an essential part of different lab-on-a-chip sample preparation methods, which aim at the release, separation, and subsequent analysis of DNA, proteins, or metabolites. Particularly for the analysis of compartmented in vivometabolism of mammalian cells, such a method must be very fast compared to the metabolic turnover-rates, it should not affect the native metabolite concentrations, and should ideally leave cell organelles undamaged. So far, no such a method is available. We have developed a microfluidic system for the effective rapid mechanical cell disruption and established a mathematical model to describe the efficiency of the system. Chinese hamster ovary (CHO) cells were disrupted with high efficiency by passing through two consecutive micronozzle arrays. Simultaneous cell compression and shearing led to a disruption rate of ≥90% at a sample flow rate of Q = 120 μL min−1 per nozzle passage, which corresponds to a mean fluid velocity of 13.3 m s−1 and a mean Reynolds number of 22.6 in the nozzle gap. We discussed the problem of channel clogging by cellular debris and the resulting flow instability at the micronozzle arrays. The experimental results were compared to predictions from Computational Fluid Dynamics (CFD) simulations and the critical energy dissipation rate for the disruption of the CHO cell population with known size distribution was determined to be 4.7 × 108 W m−3. Our model for the calculation of cell disruption on the basis of CFD-data could be applied to other microgeometries to predict intended disruption or undesired cell damage.