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Bio-Design and Manufacturing  2023 Vol.6 No.1 P.1-11

http://doi.org/10.1007/s42242-022-00205-3


Microcapillary cell extrusion deposition with picolitre dispensing resolution


Author(s):  Saeed Fathi, Iek Man Lei, Yang Cao & Yan Yan Shery Huang

Affiliation(s):  Department of Engineering, University of Cambridge, Cambridge, UK; more

Corresponding email(s):   sf629@cam.ac.uk, saeed.fathi@gmail.com

Key Words:  Microcapillary cell printing, Resolution dispensing, Cell sedimentation, Single cell


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Saeed Fathi, Iek Man Lei, Yang Cao & Yan Yan Shery Huang. Microcapillary cell extrusion deposition with picolitre dispensing resolution[J]. Journal of Zhejiang University Science D, 2023, 6(1): 1-11.

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Abstract: 
Extrusion-based cell deposition has become a prominent technique for expanding bioprinting applications. However, the associated print resolution in the order of nanolitre or above has been a limiting factor. The demand for improving print resolution towards the scale of a single cell has driven the development of precision nozzle extrusion, although the benefits gained remain ambiguous. Here, aided by in situ imaging, we investigated the dynamics of cell organisation through an extrusion-based microcapillary tip with picolitre precision through in-air or immersion deposition. The microcapillary extrusion setup, termed Picodis, was demonstrated by generating droplets of colouring inks immersed in an immiscible medium. Next, using 3T3 fibroblast cells as an experimental model, we demonstrated the deposition of cell suspension, and pre-aggregated cell pellets. Then, the dynamic organisation of cells within the microcapillary tip was described, along with cell ejection and deposition upon exiting the tip opening. The vision-assisted approach revealed that when dispersed in a culture medium, the movements of cells were distinctive based on the flow profiles and were purely driven by laminar fluid flow within a narrow tip. The primary process limitations were cell sedimentation, aggregation and compaction, along with trapped air bubbles. The use of picolitre-level resolution microcapillary extrusion, although it provides some level of control for a small number of cells, does not necessarily offer a reliable method when a specified number of cells are required. Our study provides insights into the process limitations of high-resolution cell ink extrusion, which may be useful for optimising biofabrication processes of cell-laden constructs for biomedical research.

剑桥大学Saeed Fathi等 | 具有皮升级别精度的微毛细管细胞挤出打印

本研究论文聚焦单细胞精度生物3D打印工艺技术。基于挤出式方法的细胞沉积技术已成为一种重要的技术,它可以扩展生物打印应用。然而,纳升或更高数量级的打印精度一直限制着挤出式打印。虽然所获得的好处仍不明确,将打印精度提高到单个细胞精度的需求推动了精准挤出的发展。这里,在原位成像技术的帮助下,我们通过具有皮升精度的微毛细管尖端在空气中或浸入条件下的挤出方法,探究该过程中细胞组织的动力学。我们通过在不混溶介质中的产生着色墨水液滴的方法来证明这套名为“Picodis”的微毛细管挤出装置。接下来,使用3T3成纤维细胞作为实验模型,展示了细胞悬浮液和预聚集细胞颗粒的挤出沉积。然后,描述了微毛细血管尖端内细胞的动态组织,以及在离开尖端开口时细胞的排出和沉积。通过视觉辅助方法,揭示了当细胞分散在培养基中时,其独特的基于流量剖面的运动,该运动仅仅受到狭窄尖端内的层流流动驱动。研究中,主要的工艺限制是细胞沉降、聚集和挤压,以及含有的气泡。虽然使用皮升级精度微毛细管挤出能在某种程度上控制少量细胞,但是在需要特定数量的细胞时,这种方法不一定可靠。我们的研究提供了针对高精度细胞墨水挤出时受到的工艺限制的见解,这可能有助于优化生物医学研究中对载细胞结构制造的工艺。

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