Label-free microfluidic isolation of functional and viable lymphocytes from peripheral blood mononuclear cells.
Label-free microfluidic isolation of functional and viable lymphocytes from peripheral blood mononuclear cells.
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DOI：
10.1063/5.0161047
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发表时间：
2023-09
期刊：
影响因子：
3.2
通讯作者：
Abhishek Raj;Katily Ramirez;Katherine M. Young;Nicholas E. Stone;Peter Shankles;Mehdia Nadeem Rajab Ali;Anthony Malik Compton;Wilbur A. Lam;Alexander Alexeev;Todd Sulchek
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作者：
Abhishek Raj;Katily Ramirez;Katherine M. Young;Nicholas E. Stone;Peter Shankles;Mehdia Nadeem Rajab Ali;Anthony Malik Compton;Wilbur A. Lam;Alexander Alexeev;Todd Sulchek
将外周血单个核细胞(PBMC)分离成不同类型的血细胞是获得免疫细胞进行自体细胞治疗的重要步骤。基于生物力学性质的差异，使用无标记微流控技术分离PBMC的能力与其他传统技术相比具有许多优点，包括成本更低、使用方便、避免了动物来源的标记抗体。在这里，我们报告了一种微流控装置，它使用压缩对角脊将PBMC分离成高纯度的活的和功能的淋巴细胞样本。这项技术利用PBMC亚群的生物物理特性的差异，将淋巴细胞和单核细胞引导到不同的出口。首先用原子力显微镜表征了来自健康捐献者的单核细胞和淋巴细胞的生物物理性质。淋巴细胞的硬度明显高于单核细胞，平均细胞硬度分别为1495和931pA。生物物理特性的差异导致了通过终止于不同出口的微通道的不同轨迹，导致淋巴细胞样本的纯度和活力均高于96%，而不影响细胞产生干扰素伽马的能力，干扰素伽马是一种对先天免疫和获得性免疫至关重要的细胞因子。
The separation of peripheral blood mononuclear cells (PBMCs) into constituent blood cell types is a vital step to obtain immune cells for autologous cell therapies. The ability to separate PBMCs using label-free microfluidic techniques, based on differences in biomechanical properties, can have a number of benefits over other conventional techniques, including lower cost, ease of use, and avoidance of animal-derived labeling antibodies. Here, we report a microfluidic device that uses compressive diagonal ridges to separate PBMCs into highly pure samples of viable and functional lymphocytes. The technique utilizes the differences in the biophysical properties of PBMC sub-populations to direct the lymphocytes and monocytes into separate outlets. The biophysical properties of the monocytes and lymphocytes from healthy donors were first characterized using atomic force microscopy. Lymphocytes were found to be significantly stiffer than monocytes, with a mean cell stiffness of 1495 and 931 Pa, respectively. The differences in biophysical properties resulted in distinct trajectories through the microchannel terminating at different outlets, resulting in a lymphocyte sample with purity and viability both greater than 96% with no effect on the cells' ability to produce interferon gamma, a cytokine crucial for innate and adaptive immunity.