Good manufacturing practice production of human corneal limbus-derived stromal stem cells and in vitro quality screening for therapeutic inhibition of corneal scarring

• Established a complete GMP-compliant protocol for isolation, expansion, and cryopreservation of human corneal stromal stem cells (CSSCs), addressing a critical step toward clinical-grade cell manufacturing. • Developed a novel in vitro quality control metric, the Scarring Index (SI), based on stemness markers (ABCG2, nestin) and anti-inflammatory activity, which accurately predicts in vivo anti-scarring potency. • Demonstrated that CSSC batches with SI < 10 achieve at least 50% scar reduction in a mouse corneal injury model, whereas SI > 10 indicates ineffectiveness, providing a reliable potency assay. • The GMP-compliant production and QC strategy is broadly applicable to other cell-based therapies and pharmacological treatments for corneal scarring and beyond.