Analysis of different cell culture systems for isolation, cultivation and plating of SARS-CoV-2

SARS-CoV-2 was isolated from several respiratory samples (bronchoalveolar lavage, nasal swab, oropharyngeal swab, nasopharyngeal lavage) and stool samples, in different cell lines and in several laboratories around the world such as China, Australia, USA, Germany, Brazil and South Korea. The first isolation was reported in China in the study describing the new coronavirus published on January 24, 2020 (Zhu et al., 2020). The virus was isolated from lower respiratory tract samples from four patients with pneumonia of unknown etiology, collected on December 21, 2019, all with a history of passage in the Seafood market Huanan, Wuhan, epicenter of the SARS epidemic. CoV-2. In this first report, primary culture inoculation of bronchoalveolar epithelial cells obtained from human lungs of donors was used. These cells are held in a collagen membrane at an air-liquid interface. Later reports indicated the possibility of cultivation in cell lines more commonly used as the VERO E6 and HUH-7 cells. In the present study, we intend to compare the efficacy of different cell lines in the cultivation of SARS-CoV-2 including HRT-18g, Caco2, LLC-MK2 and HUH-7 cells. We will evaluate the replicative capacity, the presence of cytopathic effect, the capacity to form plaques and the ease of visualizing them and days after inoculation for their formation.