Fig. 2: Generation of ICAM-1 Knockout Pluripotent Stem Cell (PSC) Lines.

ICAM-1 was knocked-out (KO) via CRISPR/Cas9 editing. A Schematic demonstrating the CRISPR/Cas9 KO strategy. The wild type (WT) codon (CTG) in the first exon of the ICAM-1 sequence is edited into a stop codon (TGA) via the addition of a nucleotide. Created in BioRender. Saha, S. (https://BioRender.com/slgw6qa). B Representative PSC colony morphology (10X) of > 3 independent experiments. Images were acquired on ECHO Revolve | R4 microscope. C Pluripotency marker SSEA-4 staining by flow cytometry. Unstained controls are shown with a dashed line. Data representative of 3 independent experiments. Analysis was performed via FlowJo v10 software. D Teratomas were grown in immune-deficient mouse hosts by intramuscular injection of the H9 ICAM-1 KO PSC line with Matrigel. Representative (>3 independent experiments) Hematoxylin and Eosin-stained images of (top) gut [Endoderm], (middle) cartilage [Mesoderm], and (bottom) retinal pigment epithelium [Ectoderm] representing all three germ layers. Multiple cell lines were made in human embryonic stem cells (hESCs) and in human induced PSCs; data are representative of all lines. Images were acquired on ECHO Revolve | R4 microscope.