A novel large-scale production system for modified basement membrane matrices using gene-swapped parietal endoderm cells

• DOI: 10.1016/j.matbio.2005.10.004
• Publication Date: 2005-11-29T12:47:26Z
• Authors: Yoshitaka Hayashi; Charles N. Weber; Tomomi Emoto; Hironobu Fujiwara; Noriko Sanzen; Sugiko Futaki; Kiyotoshi Sekiguchi
• Keywords: 0301 basic medicine; 03 medical and health sciences; Cell Line, Tumor; Genetic Vectors; Animals; Gene Expression Regulation, Developmental; Laminin; Basement Membrane

Parietal endoderm-like cells, including Engelbreth-Holm-Swarm tumor and differentiated F9 embryonal carcinoma cells, produce huge amounts of basement membrane components, including laminin-1 (alpha1beta1gamma1). We employed a double-lox system-based gene-swapping strategy in F9 cells to replace the laminin alpha1 gene with a laminin alpha5 minigene. The gene-swapped F9 cells secreted laminin-10 (alpha5beta1gamma1) consisting of the exogenous alpha5 subunit and endogenous beta1 and gamma1 subunits on differentiation. The laminin-10 concentration in the conditioned medium exceeded 10 mg/l, which is 10-fold higher than the concentrations achieved by conventional recombinant expression systems. The gene-swapped F9 cells deposited basement membrane-like matrices containing laminin-10 on culture dishes, offering a novel microenvironment for in vitro cell manipulation.