Multiscale Bioengineering, Faculty of Technology, Bielefeld University, Universitätsstraße 25, 33615, Bielefeld, Germany.
Center for Biotechnology (CeBiTec), Bielefeld University, Universitätsstraße 27, 33615, Bielefeld, Germany.
Sci Rep. 2023 Mar 8;13(1):3857. doi: 10.1038/s41598-023-30297-5.
Microfluidic cultivation, with its high level of environmental control and spatio-temporal resolution of cellular behavior, is a well-established tool in today's microfluidics. Yet, reliable retention of (randomly) motile cells inside designated cultivation compartments still represents a limitation, which prohibits systematic single-cell growth studies. To overcome this obstacle, current approaches rely on complex multilayer chips or on-chip valves, which makes their application for a broad community of users infeasible. Here, we present an easy-to-implement cell retention concept to withhold cells inside microfluidic cultivation chambers. By introducing a blocking structure into a cultivation chamber's entrance and nearly closing it, cells can be manually pushed into the chamber during loading procedures but are unable to leave it autonomously in subsequent long-term cultivation. CFD simulations as well as trace substance experiments confirm sufficient nutrient supply within the chamber. Through preventing recurring cell loss, growth data obtained from Chinese hamster ovary cultivation on colony level perfectly match data determined from single-cell data, which eventually allows reliable high throughput studies of single-cell growth. Due to its transferability to other chamber-based approaches, we strongly believe that our concept is also applicable for a broad range of cellular taxis studies or analyses of directed migration in basic or biomedical research.
微流控培养技术具有高度的环境控制和细胞行为的时空分辨率，是当今微流控领域中一种成熟的工具。然而，可靠地将（随机）游动细胞保留在指定的培养隔室中仍然是一个限制，这限制了系统的单细胞生长研究。为了克服这一障碍，目前的方法依赖于复杂的多层芯片或芯片阀，这使得它们的应用对于广大用户来说是不可行的。在这里，我们提出了一种易于实现的细胞保留概念，将细胞保留在微流控培养室中。通过在培养室的入口处引入一个阻塞结构并将其几乎关闭，可以在加载过程中手动将细胞推入室中，但在随后的长期培养中，细胞无法自主离开。CFD 模拟和痕量物质实验证实了室内有足够的营养供应。通过防止细胞反复丢失，从中国仓鼠卵巢培养的集落水平获得的生长数据与单细胞数据确定的数据完全匹配，最终可以可靠地进行高通量单细胞生长研究。由于其可转移到其他基于腔室的方法，我们坚信我们的概念也适用于广泛的细胞趋药性研究或基础或生物医学研究中的定向迁移分析。
