CTGF knockdown in Vero cells reduces autophagy and adhesion and promotes short-term suspension adaptation.

Introduction: Vero cells are extensively used in viral vaccine production, but their adaptation to serum-free suspension culture is hindered by excessive autophagy and strong anchorage dependence. Methods: In this study, we identified Connective Tissue Growth Factor (CTGF/CCN2) as significantly upregulated under starvation-an inducer of autophagy-via RNA-seq screening. A stable CTGF knockdown Vero cell line (knockdown efficiency >50%) was established using lentiviral shRNA. Results: Functional characterization demonstrated that CTGF depletion concurrently attenuated autophagic flux (evidenced by reduced LC3-II/I ratio and lysosomal activity) and impaired cell adhesion (with adhesion rates decreased by 50%-60% on extracellular matrix proteins), while maintaining normal cell proliferation. Discussion: Our findings reveal a new role for CTGF in regulating the environmental adaptation of Vero cells by coordinating autophagy and adhesion functions. The engineered cell line provides a novel strategy to overcome suspension adaptation bottlenecks, offering significant potential for improving vaccine production scalability.