Mahdavi Reza, Hashemi-Najafabadi Sameereh, Ghiass Mohammad Adel, Valaskivi Silmu, Välimäki Hannu, Kreutzer Joose, Hamngren Blomqvist Charlotte, Romeo Stefano, Kallio Pasi, Adiels Caroline Beck
Biotechnology Department, Faculty of Chemical Engineering, Tarbiat Modares University, Tehran, Iran.
Biomedical Engineering Department, Faculty of Chemical Engineering, Tarbiat Modares University, P.O. Box, Tehran, 14115-114, IR, Iran.
Biomed Microdevices. 2025 Feb 14;27(1):8. doi: 10.1007/s10544-025-00738-1.
Liver zonation is a fundamental characteristic of hepatocyte spatial heterogeneity, which is challenging to recapitulate in traditional cell cultures. This study presents a novel microfluidic device designed to induce zonation in liver cell cultures by establishing an oxygen gradient using standard laboratory gases. The device consists of two layers; a bottom layer containing a gas channel network that delivers high (cell incubator air, 19% oxygen) and low oxygenated (nitrogen) gases to create three distinct zones within the cell culture chamber in the layer above. Computational simulations and ratiometric oxygen sensing were employed to validate the oxygen gradient, demonstrating that stable oxygen levels were achieved within two hours. Liver zonation was confirmed using immunofluorescence staining, which showed zonated albumin production in HepG2 cells directly correlating with oxygen levels and mimicking in-vivo zonation behavior. This user-friendly device supports studies on liver zonation and related metabolic disease mechanisms in vitro. It can also be utilized for experiments that necessitate precise gas concentration gradients, such as hypoxia-related research areas focused on angiogenesis and cancer development.
肝小叶分区是肝细胞空间异质性的一个基本特征，这在传统细胞培养中很难重现。本研究提出了一种新型微流控装置，该装置旨在通过使用标准实验室气体建立氧梯度来诱导肝细胞培养中的小叶分区。该装置由两层组成；底层包含一个气体通道网络，该网络输送高氧（细胞培养箱空气，19%氧气）和低氧（氮气）气体，以在上方层的细胞培养室内创建三个不同的区域。采用计算模拟和比率氧传感来验证氧梯度，结果表明在两小时内实现了稳定的氧水平。通过免疫荧光染色证实了肝小叶分区，结果显示HepG2细胞中分区的白蛋白产生与氧水平直接相关，并模拟了体内小叶分区行为。这种用户友好型装置支持体外肝小叶分区及相关代谢疾病机制的研究。它还可用于需要精确气体浓度梯度的实验，如专注于血管生成和癌症发展的缺氧相关研究领域。
