Lee Kang Kug Paul, Matsu-Ura Toru, Rosselot Andrew E, Broda Taylor R, Wells James M, Hong Christian I
Computational and Molecular Biology Laboratory, Department of Pharmacology and Systems Physiology, University of Cincinnati, Cincinnati, Ohio 45267, USA.
Biomicrofluidics. 2021 Feb 17;15(1):014110. doi: 10.1063/5.0036527. eCollection 2021 Jan.
Perfused three-dimensional (3D) cultures enable long-term growth and monitoring of 3D organoids making them well-suited for investigating organoid development, growth, and function. One of the limitations of this long-term on-chip perfused 3D culture is unintended and disruptive air bubbles. To overcome this obstacle, we invented an imaging platform that integrates an innovative microfluidic bubble pocket for long-term perfused 3D culture of gastrointestinal (GI) organoids. We successfully applied 3D printing technology to create polymer molds that cast polydimethylsiloxane (PDMS) culture chambers in addition to bubble pockets. Our developed platform traps unintended, or induced, air bubbles in an integrated PDMS pocket chamber, where the bubbles diffuse out across the gas permeable PDMS or an outlet tube. We demonstrated that our robust platform integrated with the novel bubble pocket effectively circumvents the development of bubbles into human and mouse GI organoid cultures during long-term perfused time-course imaging. Our platform with the innovative integrated bubble pocket is ideally suited for studies requiring long-term perfusion monitoring of organ growth and morphogenesis as well as function.
灌注式三维（3D）培养能够实现3D类器官的长期生长和监测，使其非常适合用于研究类器官的发育、生长和功能。这种长期芯片上灌注式3D培养的局限性之一是意外出现且具有破坏性的气泡。为克服这一障碍，我们发明了一种成像平台，该平台集成了一个创新的微流控气泡阱，用于胃肠道（GI）类器官的长期灌注式3D培养。我们成功应用3D打印技术制作聚合物模具，除了气泡阱之外，还能铸造聚二甲基硅氧烷（PDMS）培养室。我们开发的平台将意外产生或引入的气泡捕获在集成的PDMS阱室中，气泡会通过透气的PDMS或出口管扩散出去。我们证明，在长期灌注时间进程成像过程中，我们与新型气泡阱集成的强大平台有效地避免了人类和小鼠GI类器官培养中气泡的形成。我们具有创新集成气泡阱的平台非常适合需要对器官生长、形态发生以及功能进行长期灌注监测的研究。
