Modulation of acoustofluidic parameters to assess effect on molecular loading in human T cells.
Modulation of acoustofluidic parameters to assess effect on molecular loading in human T cells.
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调节声流体参数以评估对人类 T 细胞分子负载的影响。
DOI：
10.1121/10.0007543
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发表时间：
2021
期刊：
影响因子：
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通讯作者：
Kopechek, Jonathan.
中科院分区：
文献类型：
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作者：
Centner, Connor;Moore, John;Baxter, Mary;Vinseiro Figueira, Mariana;Long, Zachary;Belott, Clinton;Menze, Michael;Bates, Paula;Yaddanapudi, Kavitha;Kopechek, Jonathan.
T 细胞疗法正在迅速兴起，用于治疗癌症和其他疾病，但受到低效的非病毒传递方法的限制。声流控装置正在开发中，以增强非病毒向细胞的传递。使用 3D 打印的声流控装置评估了声流控参数（例如通道几何形状）对人类 T 细胞分子负载的影响。将具有直线通道（直径 1 毫米和 2 毫米）的装置直接与同心螺旋通道几何形状进行比较。在用阳离子微泡 (2.5% v/v) 进行超声处理后，使用流式细胞术在 Jurkat T 细胞中评估荧光染料（钙黄绿素，100 lg/ml）的细胞内递送。 B 模式超声脉冲（2.5 MHz，3.8 MPa 输出压力）由 Verasonics Vantage 超声系统上的 P4-1 换能器生成。使用碘化丙啶染色（10 lg/ml）评估细胞活力。每个通道几何形状中的声流控处理均显着增强了细胞内分子递送，但与 1 毫米和 2 毫米直线通道相比，1 毫米同心螺旋几何形状的处理进一步增强了声流控处理后的递送（方差分析 p < 0.001，n = 6/组）。这些结果表明，3D 打印的声流体装置增强了向 T 细胞的分子递送，并且通道几何形状调节细胞内的装载效率。这种方法可能为改善 T 细胞疗法的生产提供优势。
T-cell therapies are rapidly emerging for treatment of cancer and other diseases but are limited by inefficient non-viral delivery methods. Acoustofluidic devices are in development to enhance non-viral delivery to cells. The effect of acoustofluidic parameters, such as channel geometry, on molecular loading in human T cells was assessed using 3D-printed acoustofluidic devices. Devices with rectilinear channels (1- and 2-mm diameters) were compared directly with concentric spiral channel geometries. Intracellular delivery of a fluorescent dye (calcein, 100 lg/ml) was evaluated in Jurkat T cells using flow cytometry after ultrasound treatment with cationic microbubbles (2.5% v/v). B-mode ultrasound pulses (2.5 MHz, 3.8 MPa output pressure) were generated by a P4-1 transducer on a Verasonics Vantage ultrasound system. Cell viability was assessed using propidum iodine staining (10 lg/ml). Intracellular molecular delivery was significantly enhanced with acoustofluidic treatment in each channel geometry, but treatment with the 1-mm concentric spiral geometry further enhanced delivery after acoustofluidic treatment compared to both 1- and 2-mm rectilinear channels (ANOVA p < 0.001, n ¼ 6/group). These results indicate that 3Dprinted acoustofluidic devices enhance molecular delivery to T cells, and channel geometry modulates intracellular loading efficiency. This approach may offer advantages to improve manufacturing of T cell therapies.