利用含有纳米自组装肽RADA16-I（纳米凝胶）的水凝胶作为支架，建立了气道器官样体和腺病毒感染模型。结果支持体外腺病毒研究，包括隔离和培养、发病机制研究和抗病毒药物筛选。HSAEC1-KT、HuLEC-5a和HELF细胞在RADA16-I水凝胶支架中共同培养构建了气道器官样体模型。然后使用腺病毒感染该模型进行腺病毒相关研究。评估了腺病毒感染前后气道器官样体的形态特征、增殖和活性，并检测了气道器官样体标记蛋白CC10、KRT8、AQP5、SPC、VIM和CD31的表达。采用透射电子显微镜和荧光定量PCR检测气道器官样体中腺病毒的增殖情况。 HSAEC1-KT、HuLEC-5a和HELF细胞以10:7:2的比例在RADA16-I水凝胶3D培养系统中自组装成气道器官样体，并保持长期增殖。这些器官样体稳定表达形成腔泡的蛋白KRT8和终末气道标记物AQP5和SPC。腺病毒在该模型中保持了长期增殖。 利用RADA16-I水凝胶在体外构建了由三种人类肺源性细胞系构成的气道腺病毒感染模型。该模型具有潜力作为新型腺病毒隔离和培养、发病机制研究和抗病毒药物筛选的研究工具。 © 2023 徐等

Hydrogels containing the nano-self-assembling peptide RADA16-I (Nanogels) were utilized as scaffolds to establish airway organoids and an adenovirus-infected model. The results support in vitro adenovirus studies, including isolation and culture, pathogenesis research, and antiviral drug screening.HSAEC1-KT, HuLEC-5a and HELF cells were cocultured in RADA16-I hydrogel scaffolds to construct an airway organoid model. Adenovirus was used to infect this model for adenovirus-related studies. The morphological characteristics and the proliferation and activity of airway organoids before and after adenovirus infection were evaluated. The expression of the airway organoid marker proteins CC10, KRT8, AQP5, SPC, VIM and CD31 was detected. TEM and qPCR were used to detect adenovirus proliferation in airway organoids.HSAEC1-KT, HuLEC-5a and HELF cells cocultured at 10:7:2 self-assembled into airway organoids and maintained long-term proliferation in a RADA16-I hydrogel 3D culture system. The organoids stably expressed the lumen-forming protein KRT8 and the terminal airway markers AQP5 and SPC. Adenoviruses maintained long-term proliferation in this model.An airway-organoid model of adenovirus infection was constructed in vitro from three human lung-derived cell lines on RADA16-I hydrogels. The model has potential as a novel research tool for adenovirus isolation and culture, pathogenesis research, and antiviral drug screening.© 2023 Xu et al.