Ebrahimi Aliakbar, Ghorbanpoor Hamed, Apaydın Elif, Demir Cevizlidere Bahar, Özel Ceren, Tüfekçioğlu Emre, Koç Yücel, Topal Ahmet Emin, Tomsuk Özlem, Güleç Kadri, Abdullayeva Nuran, Kaya Murat, Ghorbani Aynaz, Şengel Tayfun, Benzait Zineb, Uysal Onur, Eker Sarıboyacı Ayla, Doğan Güzel Fatma, Singh Hemant, Hassan Shabir, Ankara Hüseyin, Pat Suat, Atalay Eray, Avci Huseyin
Cellular Therapy and Stem Cell Production Application and Research Center (ESTEM), Eskisehir Osmangazi University, Eskisehir, Türkiye.
Cellular Therapy and Stem Cell Production Application and Research Center (ESTEM), Eskisehir Osmangazi University, Eskisehir, Türkiye; Department of Biomedical Engineering, Eskisehir Osmangazi University, Eskisehir, Türkiye.
Colloids Surf B Biointerfaces. 2025 Jan;245:114292. doi: 10.1016/j.colsurfb.2024.114292. Epub 2024 Oct 3.
Liver is responsible for the metabolization processes of up to 90 % of compounds and toxins in the body. Therefore liver-on-a-chip systems, as an in vitro promising cell culture platform, have great importance for fundamental science and drug development. In most of the liver-on-a-chip studies, seeding cells on both sides of a porous membrane, which represents the basement membrane, fail to resemble the native characteristics of biochemical, biophysical, and mechanical properties. In this study, polycarbonate (PC) and polyethylene terephthalate (PET) membranes were coated with gelatin to address this issue by accurately mimicking the native basement membrane present in the space of Disse. Various coating methods were used, including doctor blade, gel micro-injection, electrospinning, and spin coating. Spin coating was demonstrated to be the most effective technique owing to the ability to produce thin gel thickness with desirable surface roughness for cell interactions on both sides of the membrane. HepG2 and EA.HY926 cells were seeded on the upper and bottom sides of the gelatin-coated PET membrane and cultured on-chip for 7 days. Cell viability increased from 90 % to 95 %, while apoptotic index decreased. Albumin secretion notably rose between days 1-7 and 4-7, while GST-α secretion decreased from day 1 to day 7. In conclusion, the optimized spin coating process reported here can effectively modify the membranes to better mimic the native basement membrane niche characteristics.
肝脏负责体内高达90%的化合物和毒素的代谢过程。因此，芯片肝脏系统作为一种有前景的体外细胞培养平台，对基础科学和药物开发具有重要意义。在大多数芯片肝脏研究中，将细胞接种在代表基底膜的多孔膜两侧，无法模拟生化、生物物理和机械特性的天然特征。在本研究中，聚碳酸酯（PC）和聚对苯二甲酸乙二酯（PET）膜用明胶包被，以通过精确模拟狄氏间隙中存在的天然基底膜来解决这一问题。使用了各种包被方法，包括刮刀法、凝胶微注射法、静电纺丝法和旋涂法。旋涂法被证明是最有效的技术，因为它能够产生具有理想表面粗糙度的薄凝胶层，有利于膜两侧的细胞相互作用。将HepG2和EA.HY926细胞接种在明胶包被的PET膜的上侧和下侧，并在芯片上培养7天。细胞活力从90%提高到95%，而凋亡指数降低。白蛋白分泌在第1 - 7天和第4 - 7天显著增加，而谷胱甘肽S-转移酶α（GST-α）分泌从第1天到第7天减少。总之，本文报道的优化旋涂工艺可以有效地修饰膜，以更好地模拟天然基底膜微环境特征。
