Automated Addressable Microfluidic Device for Minimally Disruptive Manipulation of Cells and Fluids within Living Cultures
Automated Addressable Microfluidic Device for Minimally Disruptive Manipulation of Cells and Fluids within Living Cultures
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DOI：
10.1021/acsbiomaterials.9b01969
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发表时间：
2020-03-01
影响因子：
5.8
通讯作者：
Voronov, Roman
中科院分区：
文献类型：
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作者：
Anh Tong;Quang Long Pham;Voronov, Roman
细胞培养实验对于生物学研究、新医学治疗的开发和生物制造工业是普遍存在的。然而，仍然存在重大的技术障碍，限制了知识的进步，并使与这些系统相关的实验成本激增。例如，目前，难以对培养样品中的细胞进行非破坏性监测和控制。这通常需要使用牺牲测定法，并导致产品不一致。为了解决这些瓶颈，我们提出了一个原型“可寻址”的微流体技术能够在活的培养液中进行时空流体和细胞操作。作为概念验证，我们证明了其通过以空间模式接种细胞（包括共培养多种细胞类型）进行增材制造的能力，以及通过胰蛋白酶的聚焦流去除表面粘附细胞进行减材制造的能力。此外，我们表明，该设备可以从其培养室内的任何位置采样流体并进行细胞“活检”（随后可以发送进行非原位分析）。最后，芯片上的管道是完全自动化的使用外部电子设备。这开启了进行长期计算机驱动实验的可能性，其中细胞行为响应于最小破坏性观察（例如，流体取样和细胞活组织检查）。所提出的原型的局限性是它只是二维（2D）的。然而，技术是最终将概念扩展到三维（3D）的基础。该设备的另一个局限性是，它目前是由聚二甲基硅氧烷（PDMS）制成的，而需要做更多的工作来制造降解或允许细胞沉积组织基质的材料。不幸的是，现有的生物可降解材料通常不足以制造微流体阀。因此，在这项技术成为主流之前，需要开发新的技术。然而，作者希望这一目标能够很快实现，并且微流体管道技术最终将扩大到3D，以克服传统细胞培养平台的局限性。
Cell culturing experiments are ubiquitous to the study of biology, development of new medical treatments, and the biomanufacturing industry. However, there are still major technological barriers limiting the advancement of knowledge and ballooning the experimental costs associated with these systems. For example, currently, it is difficult to perform nondisruptive monitoring and control of the cells in the cultured samples. This often necessitates the use of sacrificial assays and results in product inconsistency. To resolve these bottlenecks, we present a prototype "addressable" microfluidic technology capable of spatiotemporal fluid and cell manipulations within living cultures. As a proof-of-concept, we demonstrate its ability to perform additive manufacturing by seeding cells in spatial patterns (including co-culturing multiple cell types) and subtractive manufacturing by removing surface adherent cells via the focused flow of trypsin. Additionally, we show that the device can sample fluids and perform cell "biopsies" (which can be subsequently sent for ex situ analysis), from any location within its culture chamber. Finally, the on-chip plumbing is completely automated using external electronics. This opens the possibility of performing long-term computer-driven experiments, where the cell behavior is modulated in response to the minimally disruptive observations (e.g., fluid sampling and cell biopsies) throughout the entire duration of the cultures. A limitation of the presented a prototype is that it is only two-dimensional (2D). However, technology serves as a foundation for ultimately extending the concept to three-dimensional (3D). Another limitation of the device is that it is currently made from poly(dimethylsiloxane) (PDMS), while more work needs to be done to manufacture from a material that degrades away or allow the cells to lay down the tissue matrix. Unfortunately, the existing biodegradable materials are typically not strong enough for the fabrication of microfluidic valves. Hence, new ones need to be developed before this technology can become mainstream. Yet, it is the hope of the authors that this will be achieved soon, and the microfluidic plumbing technology will eventually be scaled up to 3D, to overcome the limitations of the conventional cell culturing platforms.