CD47 Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from the Huh-7 hepatocellular carcinoma line. These cells carry disrupted CD47 alleles, creating a loss-of-function model for investigating CD47-mediated immune evasion. The polyclonal nature provides a population-level knockout representation, suitable for pooled functional studies.
The Huh-7 cell line originates from a well-differentiated hepatocellular carcinoma of a 57-year-old Japanese male. It displays epithelial morphology and is extensively used in liver cancer research, hepatic metabolism, and drug metabolism studies. As a hepatocyte-derived cancer model, Huh-7 retains liver-specific functions and supports viral replication, making it a versatile platform for hepatocellular carcinoma investigations.
CD47 functions as a ‘don’t eat me’ signal by binding SIRP?? on macrophages, which triggers phosphorylation of SHP-1 and SHP-2, inhibiting myosin-IIA accumulation and preventing phagocytosis. Its expression is regulated by HIF-1??, NF-??B, MYC, and inflammatory cytokines (TNF??, IFN??). Beyond phagocytosis, CD47 interacts with integrins (??v??3, ??IIb??3), thrombospondin-1, and VEGFR2 to modulate cell adhesion, migration, and T-cell co-stimulation. Knockout of CD47 disrupts these interactions, enhancing macrophage-mediated tumor cell clearance and promoting anti-tumor immunity.
In hepatocellular carcinoma, CD47 is often overexpressed and linked to immune escape and poor outcomes. Disrupting CD47 in Huh-7 cells provides a clinically relevant model to study the CD47-SIRP?? axis in liver cancer. The knockout unmasks tumor cells to macrophage phagocytosis, enabling evaluation of innate immune checkpoint blockade. This model also supports preclinical testing of anti-CD47 antibodies in a hepatic environment, which is essential for understanding liver-specific responses and therapeutic development.
Applications include macrophage co-culture phagocytosis assays quantified by flow cytometry or immunofluorescence, western blotting for CD47, SIRP??, SHP-1, and SHP-2, and cell adhesion and migration assays. Co-immunoprecipitation can probe protein interactions, while drug sensitivity assays assess anti-CD47 antibody efficacy. RNA-seq enables transcriptomic profiling of CD47 loss. These cells are also suitable for biomarker discovery and immunotherapeutic research. For technical inquiries, please contact Ascent Research.
