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Construction of an Engineered CHO Cell Line for Enhanced Exosome Secretion

WANG Ruping^1,2#, MAN Weiling^2#, WANG Lijuan^1#, LI Xiang², ZHU Tianhao², HE Yingying^1,2, LIN Yanli², WU Xiaojie²,CUI Yumeng², LI Yanghua², PANG Fang², LI Ying², MA Yue², ZHOU Chengjiang^1*, WANG Youliang^2*

(¹Baotou Medical College, Inner Mongolia University of Science and Technology, Baotou 014060, China;²Laboratory of Advanced Biotechnology, Academy of Military Medical Sciences, Academy of Military Sciences, Beijing 100071, China)

Abstract:

Exosomes, as natural drug delivery carriers, possess advantages such as high biocompatibility,stability, and low immunogenicity, demonstrating significant application potential in biomedical research. However,their naturally low secretion yield severely limits clinical translation. In this study, the efficient exogenous gene expression capability of CHO-K1 (Chinese hamster ovary-K1) cells was combined with RMCE (recombinase-mediatedcassette exchange) technology to achieve site-specific integration of key genes involved in exosome secretionregulation, including NADB (energy metabolism regulation), STEAP3 (vesicle formation regulation), and SDC-4(exosome secretion promotion). A stable engineered CHO-K1 tool cell line with enhanced exosome production wasthereby established. The results showed that site-specific integration of exosome enhancers had no adverse effect oncell growth. NTA (nanoparticle tracking analysis) further demonstrated that the number of exosome particles secreted by the monoclonal cell line clone1 increased by approximately fivefold compared with wild-type CHO-K1 cells.Therefore, this study successfully established an engineered exosome production system based on CHO-K1 cells to achieve stable and high-efficiency exosome secretion, laying a foundation for large-scale exosome manufacturing.

CSTR: 32200.14.cjcb.2026.06.0008