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一种用于研究细胞间通讯的快速共培养冲压装置。

A rapid co-culture stamping device for studying intercellular communication.

机构信息

School of Mechanical and Manufacturing Engineering, University of New South Wales, Sydney, NSW 2052, Australia.

Graduate School of Biomedical Engineering, University of New South Wales, Sydney, NSW 2052, Australia.

出版信息

Sci Rep. 2016 Oct 18;6:35618. doi: 10.1038/srep35618.

Abstract

Regulation of tissue development and repair depends on communication between neighbouring cells. Recent advances in cell micro-contact printing and microfluidics have facilitated the in-vitro study of homotypic and heterotypic cell-cell interaction. Nonetheless, these techniques are still complicated to perform and as a result, are seldom used by biologists. We report here development of a temporarily sealed microfluidic stamping device which utilizes a novel valve design for patterning two adherent cell lines with well-defined interlacing configurations to study cell-cell interactions. We demonstrate post-stamping cell viability of >95%, the stamping of multiple adherent cell types, and the ability to control the seeded cell density. We also show viability, proliferation and migration of cultured cells, enabling analysis of co-culture boundary conditions on cell fate. We also developed an in-vitro model of endothelial and cardiac stem cell interactions, which are thought to regulate coronary repair after myocardial injury. The stamp is fabricated using microfabrication techniques, is operated with a lab pipettor and uses very low reagent volumes of 20 μl with cell injection efficiency of >70%. This easy-to-use device provides a general strategy for micro-patterning of multiple cell types and will be important for studying cell-cell interactions in a multitude of applications.

摘要

组织发育和修复的调节依赖于相邻细胞之间的通讯。细胞微接触印刷术和微流控技术的最新进展促进了同型和异型细胞-细胞相互作用的体外研究。尽管如此,这些技术仍然很难执行,因此很少被生物学家使用。我们在这里报告了一种暂时密封的微流控印花设备的开发,该设备利用新颖的阀设计,可对两种贴壁细胞系进行图案设计,形成具有明确定义交错配置的图案,以研究细胞-细胞相互作用。我们证明了>95%的印花后细胞活力、多种贴壁细胞类型的印花以及控制接种细胞密度的能力。我们还展示了培养细胞的活力、增殖和迁移,从而能够分析细胞命运的共培养边界条件。我们还开发了一种内皮细胞和心脏干细胞相互作用的体外模型,这种相互作用被认为可以调节心肌损伤后的冠状动脉修复。该印花是使用微制造技术制造的,使用实验室移液器操作,试剂体积非常小,仅为 20 μl,细胞注射效率>70%。这种易于使用的设备为多种细胞类型的微图案化提供了一种通用策略,对于研究多种应用中的细胞-细胞相互作用非常重要。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7567/5067516/119d1896284a/srep35618-f1.jpg

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