Department of Biomedical Engineering, Case Western Reserve University, Cleveland, OH, USA.
Department of Genetics and Genome Sciences, Case Western Reserve University, Cleveland, OH, USA.
Analyst. 2022 Nov 21;147(23):5409-5418. doi: 10.1039/d2an01344d.
Paracrine signaling is challenging to study , as conventional culture tools dilute soluble factors and offer little to no spatiotemporal control over signaling. Microfluidic chips offer potential to address both of these issues. However, few solutions offer both control over onset and duration of cell-cell communication, and high throughput. We have developed a microfluidic chip designed to culture cells in adjacent chambers, separated by valves to selectively allow or prevent exchange of paracrine signals. The chip features 16 fluidic inputs and 128 individually-addressable chambers arranged in 32 sets of 4 chambers. Media can be continuously perfused or delivered by diffusion, which we model under different culture conditions to ensure normal cell viability. Immunocytochemistry assays can be performed in the chip, which we modeled and fine-tuned to reduce total assay time to 1 h. Finally, we validate the use of the chip for co-culture studies by showing that HEK293Ta cells respond to signals secreted by RAW 264.7 immune cells in adjacent chambers, only when the valve between the chambers is opened.
旁分泌信号传递难以研究，因为常规的培养工具会稀释可溶性因子，而且对信号传递几乎没有时空控制。微流控芯片提供了解决这两个问题的潜力。然而，很少有解决方案能够同时控制细胞间通讯的开始和持续时间，并实现高通量。我们开发了一种微流控芯片，用于在相邻腔室中培养细胞，腔室之间用阀门隔开，以选择性地允许或阻止旁分泌信号的交换。该芯片具有 16 个流体输入和 128 个可单独寻址的腔室，分为 32 组 4 个腔室。可以连续灌注或扩散介质，我们根据不同的培养条件进行建模，以确保细胞正常存活。可以在芯片中进行免疫细胞化学检测，我们对其进行了建模和微调，将总检测时间缩短到 1 小时。最后，我们通过证明当腔室之间的阀门打开时，HEK293Ta 细胞仅对相邻腔室中 RAW 264.7 免疫细胞分泌的信号作出反应，验证了该芯片在共培养研究中的应用。
