通过流动腔实验测量HL-60细胞微绒毛弹性
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国家自然科学基金资助项目(10772069, 10972081),广东省工业攻关项目(2008B011000017)


Measuring the elasticity of HL-60 cell microvillus by parallel plate flow chamber
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    摘要:

    目的 滤除平行平板流动腔实验数据中的噪声,以清晰观测细胞滚动黏附过程,并发展基于流动腔实验的细胞微绒毛弹性的测定方法。方法 运用小波分析方法技术,分离从流动腔实验中得到的E-选择素介导的HL-60细胞滚动黏附实验数据的高频热噪声信号;利用能量均分定理和拴缚细胞的力平衡方程,构建细胞微绒毛的伸长与弹性间的关系。结果 通过小波分析滤噪,可以清晰定位细胞滚动黏附的不同阶段:自由滚动-减速-停止-加速;在壁面剪切应力为0.01~0.06 Pa的条件下,源于热噪声的细胞脉动能量约占总的脉动能的80%,测得HL60细胞微绒毛的弹簧常数为(13.7±7.4) μN/m。结论 小波分析技术可滤除流动腔细胞滚动黏附实验数据中的热噪声;细胞热脉动信号中含有细胞微绒毛弹性的信息,使得人们可以采用流动腔实验技术来测量细胞微绒毛的弹性。

    Abstract:

    Objective To filter the noises in the experimental data of parallel plate flow chamber for observing more clearly the events occurring in the process of cell rolling adhesion and develop a new method to measure the elasticity of microvillus on cells based on the flow chamber experiment. Method The experiment of E-selectin regulated HL-60 cell rolling was performed by flow chamber system, and the data were denoised by wavelet analysis so that the high frequency thermal response signals were extracted from the data. Based on the equipartition theorem and equilibrium equations of tethered cell, the relationship between the cell microvillus spring constant and thermal fluctuations was constructed. Results Filtering noises from cell rolling time course by wavelet analysis, the events such as free rolling, slowing down, stopping and speeding up of rolling cell could be observed more easily; almost 80% of fluctuating energy of a rolling cell was involved in its high frequency fluctuation which was regarded as the thermal response of the cell to the Brown movement of water molecules, and the spring constant of microvillus on HL-60 cell was measured to be (13.7±7.4) μN/m at wall shear stress from 0.01~0.06 Pa. Conclusions The wavelet analysis can filter the thermal noises in cell rolling data of flow chamber experiment, and since the rigidity information of cell microvillus is involved in and can be extracted from the high frequency thermal fluctuation of the rolling cell, the parallel plate flow chamber experimental technique can be extended to measure the elasticity of microvillus on cells.

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黄强,凌颖琛,李趣欢,吴建华,方颖.通过流动腔实验测量HL-60细胞微绒毛弹性[J].医用生物力学,2010,25(4):277-282

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  • 收稿日期:2010-06-21
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