Cryogel-Integrated Biochip for Liver Tissue Engineering
Cryogel-Integrated Biochip for Liver Tissue Engineering
复制标题
DOI：
10.1021/acsabm.1c00425
复制
发表时间：
2021-07-08
影响因子：
4.7
通讯作者：
Legallais, Cecile
中科院分区：
文献类型：
--
作者：
Boulais, Lilandra;Jellali, Rachid;Legallais, Cecile
微流控系统和聚合物水凝胶已广泛应用于组织工程领域。然而，只有少数几种结合了这两种方法的工具，特别是在体外肝脏模型中，正在探索中。在这项研究中，设计了一种基于海藻酸盐的冷冻凝胶集成生物芯片，用于三维动态培养肝癌细胞。将藻酸盐冷冻凝胶在零下温度(T<；0℃)下在生物芯片中进行共价交联，以创建具有高机械稳定性和相互连接的大孔网络的支架。通过改变海藻酸盐浓度和交联剂比例，可以根据肝脏不同生理状态的硬度范围，在1.5到29 kpa之间微调冷冻胶的杨氏模数。我们证明，在该装置中，HepG2/C3A细胞可以在动态条件下培养并维持存活长达6天。随着细胞培养天数的增加，白蛋白合成和葡萄糖消耗增加。此外，在生物芯片的整个高度上观察到了3D细胞结构，该芯片在海藻酸盐裂解酶处理后被保存下来，以移除基于冷冻胶的支架。总之，这些结果证明了一种有趣的细胞培养技术的概念，该技术应该进一步研究，以设计出健康和肝硬变的肝脏模型。
Microfluidic systems and polymer hydrogels have been widely developed for tissue engineering. Yet, only a few tools combining both approaches, especially for in vitro liver models, are being explored. In this study, an alginate-based cryogel-integrated biochip was engineered for dynamic hepatoma cell line culture in three dimensions (3D). The alginate cryogel was covalently cross-linked in the biochip at subzero temperatures (T < 0 degrees C) to create a scaffold with high mechanical stability and an interconnected macroporous network. By varying the alginate concentration and the cross-linker ratio, Young's modulus of the cryogel can be fine-tuned between 1.5 and 29 kPa, corresponding to the range of stiffness of the different physiological states of the liver. We demonstrated that HepG2/C3A cells can be cultured and maintained as viable under dynamic conditions in this device up to 6 days. Albumin synthesis and glucose consumption increased over the cell culture days. Moreover, a 3D cell structure was observed across the entire height of the biochip, which was preserved following alginate lyase treatment to remove the cryogel-based scaffold. In summary, these results represent a proof of concept of an interesting cell culture technology that should be further investigated to engineer healthy and cirrhotic liver models.