Yang Xuan, Wen Xiaowei, Dai Jie, Chen Yanming, Ding Wanchuan, Wang Jun, Gu Xiang, Zhang Xuejin, Chen Jin, Sutliff Roy L, Emory Steven R, Ruan Gang
Department of Biomedical Engineering, College of Engineering and Applied Sciences, Nanjing University, Nanjing, China 210023.
Wisdom Lake Academy of Pharmacy, Xi'an Jiaotong-Liverpool University, Suzhou, China 215123.
ACS Nano. 2022 Jun 28;16(6):8751-8765. doi: 10.1021/acsnano.1c07648. Epub 2022 May 17.
Hard-to-transfect cells are cells that are known to present special difficulties in intracellular delivery of exogenous entities. However, the special transport behaviors underlying the special delivery problem in these cells have so far not been examined carefully. Here, we combine single-particle motion analysis, cell biology studies, and mathematical modeling to investigate nanoparticle transport in bone marrow-derived mesenchymal stem cells (BMSCs), a technologically important type of hard-to-transfect cells. Tat peptide-conjugated quantum dots (QDs-Tat) were used as the model nanoparticles. Two different yet complementary single-particle methods, namely, pair-correlation function and single-particle tracking, were conducted on the same cell samples and on the same viewing stage of a confocal microscope. Our results reveal significant differences in each individual step of transport of QDs-Tat in BMSCs a commonly used model cell line, HeLa cells. Single-particle motion analysis demonstrates that vesicle escape and cytoplasmic diffusion are dramatically more difficult in BMSCs than in HeLa cells. Cell biology studies show that BMSCs use different biological pathways for the cellular uptake, vesicular transport, and exocytosis of QDs-Tat than HeLa cells. A reaction-diffusion-advection model is employed to mathematically integrate the individual steps of cellular transport and can be used to predict and design nanoparticle delivery in BMSCs. This work provides dissective, quantitative, and mechanistic understandings of nanoparticle transport in BMSCs. The investigative methods described in this work can help to guide the tailored design of nanoparticle-based delivery in specific types and subtypes of hard-to-transfect cells.
难转染细胞是指那些已知在细胞内递送外源物质时存在特殊困难的细胞。然而，迄今为止，这些细胞中特殊递送问题背后的特殊转运行为尚未得到仔细研究。在这里，我们结合单粒子运动分析、细胞生物学研究和数学建模，来研究骨髓间充质干细胞（BMSC）中的纳米颗粒转运，BMSC是一类在技术上很重要的难转染细胞。将与Tat肽偶联的量子点（QDs-Tat）用作模型纳米颗粒。在共聚焦显微镜的同一细胞样本和同一观察阶段，采用了两种不同但互补的单粒子方法，即对关联函数和单粒子追踪。我们的结果揭示了QDs-Tat在BMSC（一种常用的模型细胞系）和HeLa细胞中的每一个转运步骤都存在显著差异。单粒子运动分析表明，BMSC中的囊泡逃逸和细胞质扩散比HeLa细胞困难得多。细胞生物学研究表明，与HeLa细胞相比，BMSC在QDs-Tat的细胞摄取、囊泡运输和胞吐作用方面使用不同的生物学途径。采用反应-扩散-平流模型对细胞转运的各个步骤进行数学整合，可用于预测和设计BMSC中的纳米颗粒递送。这项工作提供了对BMSC中纳米颗粒转运的剖析性、定量性和机理性理解。本文所述的研究方法有助于指导针对难转染细胞的特定类型和亚型进行基于纳米颗粒递送的定制设计。
