Injection molded open microfluidic well plate inserts for user-friendly coculture and microscopy.,Lab on a Chip

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Injection molded open microfluidic well plate inserts for user-friendly coculture and microscopy.
Lab on a Chip ( IF 6.1 ) Pub Date : 2019-11-12 , DOI: 10.1039/c9lc00706g
John H Day ₁ , Tristan M Nicholson ₂ , Xiaojing Su ₁ , Tammi L van Neel ₁ , Ivor Clinton ₁ , Anbarasi Kothandapani ₃ , Jinwoo Lee ₄ , Max H Greenberg ₅ , John K Amory ₆ , Thomas J Walsh ₇ , Charles H Muller ₈ , Omar E Franco ₅ , Colin R Jefcoate ₉ , Susan E Crawford ₅ , Joan S Jorgensen ₃ , Ashleigh B Theberge ₂

Affiliation

Department of Chemistry, University of Washington, Box 351700, Seattle, WA 98195, USA.
Department of Chemistry, University of Washington, Box 351700, Seattle, WA 98195, USA. and Department of Urology, University of Washington School of Medicine, Seattle, WA 98195, USA.
Department of Comparative Biosciences, University of Wisconsin-Madison, Madison, WI 53706, USA.
Department of Comparative Biosciences, University of Wisconsin-Madison, Madison, WI 53706, USA and Department of Cell and Regenerative Biology, University of Wisconsin-Madison, Madison, WI 53706, USA.
Department of Surgery, NorthShore University Research Institute, Affiliate of University of Chicago Pritzker School of Medicine, Evanston, IL 60201, USA.
Department of Medicine, University of Washington, Seattle, WA 98195, USA.
Department of Urology, University of Washington School of Medicine, Seattle, WA 98195, USA.
Department of Urology, University of Washington School of Medicine, Seattle, WA 98195, USA and Male Fertility Laboratory, Department of Urology, University of Washington School of Medicine, Seattle, WA 98195, USA.
Department of Cell and Regenerative Biology, University of Wisconsin-Madison, Madison, WI 53706, USA.

Open microfluidic cell culture systems are powerful tools for interrogating biological mechanisms. We have previously presented a microscale cell culture system, based on spontaneous capillary flow of biocompatible hydrogels, that is integrated into a standard cell culture well plate, with flexible cell compartment geometries and easy pipet access. Here, we present two new injection molded open microfluidic devices that also easily insert into standard cell culture well plates and standard culture workflows, allowing seamless adoption by biomedical researchers. These platforms allow culture and study of soluble factor communication among multiple cell types, and the microscale dimensions are well-suited for rare primary cells. Unique advances include optimized evaporation control within the well, manufacture with reproducible and cost-effective rapid injection molding, and compatibility with sample preparation workflows for high resolution microscopy (following well-established coverslip mounting procedures). In this work, we present several use cases that highlight the usability and widespread utility of our platform including culture of limited primary testis cells from surgical patients, microscopy readouts including immunocytochemistry and single molecule fluorescence in situ hybridization (smFISH), and coculture to study interactions between adipocytes and prostate cancer cells.

中文翻译：

注射成型的开放式微流孔板插入件，用于用户友好的共培养和显微镜检查。

开放式微流体细胞培养系统是用于研究生物学机制的强大工具。我们先前已经提出了一种基于生物相容性水凝胶的自发毛细管流的微型细胞培养系统，该系统已集成到标准细胞培养孔板中，具有灵活的细胞室几何形状和易于移液的通道。在这里，我们介绍了两种新型的注模开放式微流控设备，它们也可以轻松插入标准细胞培养孔板和标准培养工作流程中，从而使生物医学研究人员可以无缝地采用它们。这些平台允许培养和研究多种细胞类型之间的可溶性因子通讯，并且微尺度尺寸非常适合稀有原代细胞。独特的进步包括优化的井内蒸发控制，生产具有可复制且具有成本效益的快速注塑成型，并且与用于高分辨率显微镜的样品制备工作流程兼容（遵循完善的盖玻片安装程序）。在这项工作中，我们提出了几个用例，以突出我们平台的可用性和广泛用途，包括培养来自手术患者的有限的原代睾丸细胞，显微镜读数，包括免疫细胞化学和单分子荧光原位杂交（smFISH）和共培养以研究脂肪细胞与前列腺癌细胞之间的相互作用。

更新日期：2019-11-12

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