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Development of Immortalized Human Cell Lines from TERT-Transfected Multilineage Progenitor Cells (MLPC) by Differentiation or Cell Fusion: Models forSARS-CoV-2 Binding and Infectivity

Daniel P Collins, Clifford J Steer

MLPC are human cord blood-derived non-hematopoietic stem cells with extensive proliferative and differentiation capacity. TERT transfection of MLPC rendered them functionally immortal. Methods to create immortalized fully mature cells by differentiation or by direct fusion of TERT-MLPC to primary cells are described. Hepatocyte-like cells (HLC) created by differentiation and fusion displayed markers and functionality consistent with well-differentiated human hepatocytes. Differentiation of MLPC into alveolar type 2-like (AT2) cells displayed markers consistent with human primary small airway epithelial cells. Potential interactions of the SARS-CoV-2 with both cell lines were investigated by binding of spike proteins and their inhibition by spike-specific and marker-specific antibodies. Spike and spike 1 binding with AT2 cells occurred via surface ACE-2 receptors. Spike binding to both HLC and primary human hepatocytes (PHH) was mediated by interactions with the hepatocyte surface membrane asiaglycoprotein receptor 1 (ASGr1).