title: 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.affiliation: Sekiguchi Biomatrix Signaling Project, ERATO, Japanese Science and Technology Agency (JST), Aichi Medical University, 21 Karimata, Aichi-gun, Aichi 480-1195, Japan. hayashiy@riem.nagoya-u.ac.jp
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Hayashi
authors.forename: Charles N
authors.initials: CN
authors.lastname: Weber
authors.forename: Tomomi
authors.initials: T
authors.lastname: Emoto
authors.forename: Hironobu
authors.initials: H
authors.lastname: Fujiwara
authors.forename: Noriko
authors.initials: N
authors.lastname: Sanzen
authors.forename: Sugiko
authors.initials: S
authors.lastname: Futaki
authors.forename: Kiyotoshi
authors.initials: K
authors.lastname: Sekiguchi
mesh_terms: D000818:Animals; D001485:Basement Membrane / Q000196:embryology / Q000502:physiology* / Q000648:ultrastructure; D045744:Cell Line, Tumor; D018507:Gene Expression Regulation, Developmental; D005822:Genetic Vectors; D007797:Laminin / Q000235:genetics*
chemical_list: D007797:Laminin