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

Abstract

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.

• Authors: Yoshitaka Hayashi, Charles N Weber, Tomomi Emoto, Hironobu Fujiwara, Noriko Sanzen, Sugiko Futaki, Kiyotoshi Sekiguchi
• Journal: Matrix biology : journal of the International Society for Matrix Biology (Matrix Biol) Vol. 25 Issue 2 Pg. 85-8 (Mar 2006) ISSN: 0945-053X [Print] Netherlands
• PMID: 16314080 (Publication Type: Journal Article)

Chemical References

• Laminin

Topics

• Animals
• Basement Membrane (embryology, physiology, ultrastructure)
• Cell Line, Tumor
• Gene Expression Regulation, Developmental
• Genetic Vectors
• Laminin (genetics)