Raj Abhishek, Ramirez Katily, Young Katherine M, Stone Nicholas, Shankles Peter, Ali Mehdia Nadeem Rajab, Compton Anthony Malik, Lam Wilbur, Alexeev Alexander, Sulchek Todd
George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, 801 Ferst Drive, Atlanta, Georgia 30332-0405, USA.
School of Chemistry & Biochemistry, Georgia Institute of Technology, 901 Atlantic Drive, Atlanta, Georgia 30332-0400, USA.
Biomicrofluidics. 2023 Sep 18;17(5):054102. doi: 10.1063/5.0161047. eCollection 2023 Sep.
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.
将外周血单个核细胞（PBMC）分离成不同的血细胞类型是获取用于自体细胞治疗的免疫细胞的关键步骤。基于生物力学特性的差异，使用无标记微流控技术分离PBMC的能力相对于其他传统技术具有诸多优势，包括成本更低、使用方便以及避免使用动物源性标记抗体。在此，我们报道一种微流控装置，该装置利用压缩对角脊将PBMC分离成高纯度的有活力且功能正常的淋巴细胞样本。该技术利用PBMC亚群生物物理特性的差异，将淋巴细胞和单核细胞引导至不同的出口。首先使用原子力显微镜对健康供体的单核细胞和淋巴细胞的生物物理特性进行了表征。发现淋巴细胞比单核细胞明显更硬，平均细胞硬度分别为1495和931帕斯卡。生物物理特性的差异导致细胞在微通道中沿着不同轨迹行进并在不同出口终止，从而得到纯度和活力均大于96%的淋巴细胞样本，且对细胞产生γ干扰素的能力没有影响，γ干扰素是一种对固有免疫和适应性免疫至关重要的细胞因子。
