Mechanical disruption of mammalian cells in a microfluidic system and its numerical analysis based on computational fluid dynamics.
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 vivo metabolism 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 × 10(8) 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.
Hamburg University of Technology, Institute of Bioprocess and Biosystems Engineering, Hamburg, Germany.
Matthias
M
Wurm
An-Ping
AP
Zeng
D000818:Animals; D016466:CHO Cells; D002458:Cell Fractionation / Q000295:instrumentation*; D003198:Computer Simulation; D006224:Cricetinae; D003412:Cricetulus; D004867:Equipment Design; D057446:Hydrodynamics; D044085:Microfluidics / Q000295:instrumentation*; D008954:Models, Biological; D011312:Pressure; D013314:Stress, Mechanical