École Normale Supérieure-PSL Research University, Département de Chimie, Sorbonne Universités-UPMC Univ Paris 06, CNRS UMR 8640, PASTEUR, 24, Rue Lhomond, 75005, Paris, France.
MesoBioTech, 231 Rue Saint-Honoré, 75001, Paris, France.
Biosens Bioelectron. 2023 Sep 1;235:115324. doi: 10.1016/j.bios.2023.115324. Epub 2023 May 10.
Recent advances have shown the high sensibility of electrochemical impedance spectroscopy in real-time monitoring of cell barriers on a chip. Here, we applied this method to the investigation of human induced pluripotent stem cell (hiPSC) derived and artificial basement membrane (ABM) supported endothelial barrier. The ABM was obtained by self-assembling type IV collagen and laminin with a monolayer of crosslinked gelatin nanofibers. The hiPSCs were differentiated into brain microvascular endothelial cells (BMECs) and then plated on the ABM. After incubation for two days, the ABM-BMEC assembly was placed as a tissue insert into a microfluidic device for culture and real-time impedance monitoring over days. We found a significantly enhanced stability of the BMEC barrier in a serum-free and bromodeoxyuridine (BrdU) containing culture medium compared to the conventional culture due to the restricted cell proliferation. We also found that the BMEC barrier was sensitive to stimuli such as thrombin and that the change of the barrier impedance was mainly due to the change of the cell layer resistance. We can thus advocate this method to investigate the integrity of the cell barrier and the barrier-based assays.
最近的进展表明，电化学阻抗谱在实时监测芯片上的细胞屏障方面具有很高的灵敏度。在这里，我们将这种方法应用于人类诱导多能干细胞（hiPSC）衍生的和人工基底膜（ABM）支持的内皮屏障的研究。ABM 是通过自组装 IV 型胶原和层粘连蛋白与交联明胶纳米纤维单层获得的。hiPSCs 分化为脑微血管内皮细胞（BMECs），然后接种在 ABM 上。孵育两天后，将 ABM-BMEC 组件作为组织插入物放入微流控装置中进行培养和实时阻抗监测数天。我们发现，由于细胞增殖受到限制，无血清和溴脱氧尿苷（BrdU）的培养基中 BMEC 屏障的稳定性明显增强，与传统培养相比。我们还发现，BMEC 屏障对凝血酶等刺激很敏感，并且屏障阻抗的变化主要是由于细胞层电阻的变化。因此，我们可以提倡这种方法来研究细胞屏障的完整性和基于屏障的测定。
