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. 2021 Feb 13;16(3):478–492. doi: 10.1016/j.stemcr.2021.02.008

Figure 2.

Figure 2

hPSC-CMs Are Permissive to SARS-CoV-2 Infection and Replication

(A) Cytopathic effects of SARS-CoV-2 at 5 MOI in H7 hESC-CMs and WTC11c hiPSC-CMs during a time course of 120 h. Scale bars, 100 μm.

(B) Immunofluorescent staining of H7 hESC-CMs and WTC11c hiPSC-CMs at 48 hpi with SARS-CoV-2 dose of 0.1 MOI. Scale bars, 50 μm. Individual channels are shown in Figure S2C.

(C) One-step viral growth curve in H7 hESC-CMs infected with SARS-CoV-2 at 5 MOI over a time course of 120 h.

(D) Multi-step viral growth curve in H7 hESC-CMs infected with SARS-CoV-2 at 0.1 MOI over a time course of 72 h. For both (C) and (D), lines connect the means of two independent experiments. Viral RNA indicating intracellular viral replication is plotted on the left y axis as percent of HPRT1. Viral particles secreted in the supernatant are plotted on the right y axis as plaque-forming units (PFU) per mL.

(E) Cytopathic effects of 0.1 MOI and 5 MOI of SARS-CoV-2 at 72 hpi in wild-type (WT) and ACE2 knockout (KO) WTC11c-CMs. Two ACE2 KO hiPSC clones (cl.1 and cl. 2) were analyzed (Figure S3). Scale bars, 100 μm.

(F) Fluorescence microscopy assessment of viral entry of icSARS-CoV2-mNG at 0.1 MOI in WTC11c-CMs WT and ACE2 KO clones at 48 and 72 hpi. mNG fluorescence is shown in grayscale. ACE2 KO clones showed background autofluorescence only. Scale bars, 100 μm.