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

## TL;DR

Reducing CTGF in Vero cells lowers autophagy and adhesion, helping them grow better in suspension, which is important for vaccine production.

## Contribution

Identifies CTGF as a key regulator of autophagy and adhesion in Vero cells, enabling improved suspension culture for vaccine production.

## Key findings

- CTGF knockdown in Vero cells reduces autophagic flux and lysosomal activity.
- CTGF depletion impairs cell adhesion to extracellular matrix proteins by 50–60%.
- Engineered CTGF-knockdown Vero cells maintain normal proliferation while adapting better to suspension culture.

## Abstract

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.

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.

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.

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.