Purpose: To construct a three-dimensional (3D) culture model of adenovirus in vitro using the nanoself-assembling peptide RADA16-I as a 3D cell culture scaffold combined with virology experimental technology to provide a novel research method for virus isolation and culture, pathogenesis research, antiviral drug screening and vaccine preparation.
Methods: The nanoself-assembling peptide RADA16-I was used as a 3D scaffold material for 293T cell culture, and adenovirus was cultured in the cells. The growth, morphological characteristics and pathological effects of 3D-cultured 293T cells after adenovirus infection were observed with an inverted microscope and MTS. The proliferation of adenovirus in 293T cells was observed by TEM and detected by qPCR. The levels of TNF-α and IL-8 secreted by adenovirus-infected 293T cells in the RADA16-I 3D culture system were detected by ELISA.
Results: The 293T cells grew well in the RADA16-I 3D culture system for a prolonged period of time. The adenovirus infection persisted for a long time with multiple proliferation peaks, which closely resembled those of in vivo infections. The adenovirus virions amplified in the 3D system remained infectious. There were multiple secretion peaks of TNF-α and IL-8 secretion levels in adenovirus-infected 293T cells cultured in 3D culture systems.
Conclusion: The nanoself-assembling peptide RADA16-I can be used as a 3D scaffold for adenovirus isolation, culture and research. The 3D culture system shows more realistic in vivo effects than two-dimensional (2D) culture.

Keywords: nanoself-assembling peptide, 3D culture, adenovirus, 293T cells


中文翻译：

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纳米自组装肽作为3D培养支架的体外腺病毒感染研究。

目的：以纳米自组装肽RADA16-I为3D细胞培养支架，结合病毒学实验技术构建腺病毒体外三维（3D）培养模型，为病毒分离培养、发病机制提供新的研究方法。研究、抗病毒药物筛选和疫苗制备。
方法：将纳米自组装肽RADA16-I作为293T细胞培养的3D支架材料，在细胞内培养腺病毒。用倒置显微镜和MTS观察3D培养的293T细胞在腺病毒感染后的生长、形态特征和病理影响。TEM观察293T细胞中腺病毒的增殖情况，qPCR检测。ELISA检测腺病毒感染的293T细胞在RADA16-I 3D培养系统中分泌的TNF-α和IL-8水平。
结果：293T 细胞在 RADA16-I 3D 培养系统中长时间生长良好。腺病毒感染持续时间较长，具有多个增殖峰，与体内感染非常相似。在 3D 系统中扩增的腺病毒病毒粒子仍然具有传染性。在 3D 培养系统中培养的腺病毒感染的 293T 细胞中存在多个 TNF-α 和 IL-8 分泌水平的分泌峰。
结论：纳米自组装肽RADA16-I可作为腺病毒分离、培养和研究的3D支架。3D 培养系统显示出比二维 (2D) 培养更逼真的体内效果。

关键词：纳米自组装肽，3D培养，腺病毒，293T细胞