Department of Biomedical Engineering, University of California Irvine, 3107 Natural Sciences II, Irvine, CA 92697, USA.
Lab Chip. 2017 Sep 26;17(19):3300-3309. doi: 10.1039/c7lc00575j.
The ability to harvest single cells from tissues is currently a bottleneck for cell-based diagnostic technologies, and remains crucial in the fields of tissue engineering and regenerative medicine. Tissues are typically broken down using proteolytic digestion and various mechanical treatments, but success has been limited due to long processing times, low yield, and high manual labor burden. Here, we present a novel microfluidic device that utilizes precision fluid flows to improve the speed and efficiency of tissue digestion. The microfluidic channels were designed to apply hydrodynamic shear forces at discrete locations on tissue specimens up to 1 cm in length and 1 mm in diameter, thereby accelerating digestion through hydrodynamic shear forces and improved enzyme-tissue contact. We show using animal organs that our digestion device with hydro-mincing capabilities was superior to conventional scalpel mincing and digestion based on recovery of DNA and viable single cells. Thus, our microfluidic digestion device can eliminate or reduce the need to mince tissue samples with a scalpel, while reducing sample processing time and preserving cell viability. Another advantage is that downstream microfluidic operations could be integrated to enable advanced cell processing and analysis capabilities. We envision our novel device being used in research and clinical settings to promote single cell-based analysis technologies, as well as to isolate primary, progenitor, and stem cells for use in the fields of tissue engineering and regenerative medicine.
从组织中提取单细胞的能力目前是基于细胞的诊断技术的瓶颈,在组织工程和再生医学领域仍然至关重要。组织通常使用蛋白水解消化和各种机械处理来分解,但由于处理时间长、产量低和人工劳动负担高,成功的机会有限。在这里,我们提出了一种新颖的微流控装置,该装置利用精密的流体流动来提高组织消化的速度和效率。微流道的设计目的是在长达 1 厘米、直径 1 毫米的组织标本的离散位置施加流体剪切力,从而通过流体剪切力和改善的酶-组织接触来加速消化。我们通过动物器官证明,我们具有水力粉碎功能的消化装置在 DNA 和活的单细胞回收率方面优于传统的手术刀切碎和消化。因此,我们的微流控消化装置可以消除或减少使用手术刀切碎组织样本的需要,同时减少样本处理时间并保持细胞活力。另一个优点是可以集成下游微流控操作,以实现先进的细胞处理和分析功能。我们设想我们的新型设备将在研究和临床环境中使用,以促进基于单细胞的分析技术,并分离用于组织工程和再生医学领域的原代、祖细胞和干细胞。