COVID-19: 3D Bioprinted Tissue Models for Respiratory Viruses

过去几个月来，我们在实现上述若干目标方面取得了重大进展。成绩是根据目标数来写的。 1.我们已经建立了优化的方案，培养原代人小气道上皮细胞和肺泡细胞在transwell气液界面（ALI）模型。我们正在采用96孔板格式，并优化血管单元的添加。我们还获得了MatTek气管支气管和肺泡培养的丰富经验。 2.我们已经建立了人三维肺组织（气管支气管，肺泡和小气道）与流感病毒的感染和优化的检测条件。我们还建立了SARS-CoV 2感染3D ALI气管支气管组织，并能够直接和鲁棒地检测组织中的SARS-CoV 2抗原。 3.我们已经证实了Remdesivir和I型干扰素在感染SARS-CoV 2的3D ALI气管支气管组织中的强大抗病毒活性。我们正在使用这个模型来分析从高通量筛选中鉴定的选定化合物的抗病毒活性，该筛选是与宾夕法尼亚大学的Sara Cherry合作进行的。我们正在继续研究这些组织中感染的再现性，以用于筛选目的。 4. scRNAseq研究目前正在进行中。

We have made significant progress on several of our goals and objectives listed above over the past few months. Accomplishments are written according to objective number. 1. We have established optimized protocols for culturing primary human small airway epithelial cells and alveolar cells in a transwell air-liquid-interface (ALI) model. We are miniaturizing to 96-well plate formats and optimizing the addition of vasculature units. We have also gained extensive experience working with MatTek's tracheobronchial and alveolar cultures. 2. We have established infection of human 3D lung tissue (tracheobronchial, alveolar, and small airway) with influenza virus and optimized detection conditions. We have also established infection of 3D ALI tracheobronchial tissues with SARS-CoV2, and are able to directly and robustly detect SARS-CoV2 antigen in tissue. 3. We have confirmed robust antiviral activity of remdesivir and interferon type I in 3D ALI tracheobronchial tissues infected with SARS-CoV2. We are using this model to analyze the antiviral activity of select compounds identified from high throughput screening that was carried out in collaboration with Sara Cherry at the University of Pennsylvania. We are continuing to study the reproducibility of infection in these tissues for screening purposes. 4. scRNAseq studies are currently ongoing.