title: Generation of Mouse STO Feeder Cell Lines That Confer Resistance to Several Types of Selective Drugs.
abstract: Feeder cells are generally required for establishment and maintenance of embryonic stem (ES)/induced pluripotent stem (iPS) cells. Increased demands for generation of those cells carrying various types of vectors (i.e., KO vectors and transgenes) also require feeder cells that confer resistance to any types of preexisting selective drugs. Unfortunately, the use of the feeders that are resistant to various drugs appears to be limited to a few laboratories. Here we generated a set of gene-engineered STO feeder cells that confer resistance to several commercially available drugs. The STO cells, which have long been used as a feeder for mouse ES and embryonal carcinoma (EC) cells, were transfected with pcBIH [carrying bleomycin resistance gene (ble) and hygromycin B phosphotransferase gene (Hyg)], pcBIP [carryingbleand puromycin resistance gene (puro)], or pcBSN [carryingbleand neomycin resistance gene (neo)]. The resulting stably transfectants (termed SHB for pcBIH, SPB for pcBIP, and SNB for pcBSN) exhibited bleomycin/hygromycin, bleomycin/puromycin, or bleomycin/neomycin, as expected. The morphology of these cells passaged over 18 generations was indistinguishable from that of parental STO cells. Of isolated clones, the SHB3, SPB3, and SNB2 clones successfully supported the growth of mouse ES cells in an undifferentiated state, when coculture was performed. PCR analysis revealed the presence of the selective markers in these clones, as expected. These SHB3, SPB3, and SNB2 cells will thus be useful for the acquisition and maintenance of genetically manipulated ES/iPS cells.
authors.affiliation: Department of Pediatric Dentistry, Kagoshima University Graduate School of Medical and Dental Sciences , Sakuragaoka, Kagoshimashi, Kagoshima , Japan.
authors.forename: Issei
authors.initials: I
authors.lastname: Saitoh
authors.affiliation: † Section of Gene Expression Regulation, Frontier Science Research Center, Kagoshima University , Korimoto, Kagoshima, Kagoshima , Japan.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Sato
authors.affiliation: ‡ Department of Dental Anesthesia, Kagoshima University Medical and Dental Hospital , Sakuragaoka, Kagoshimashi, Kagoshima , Japan.
authors.forename: Yoko
authors.initials: Y
authors.lastname: Iwase
authors.affiliation: Department of Pediatric Dentistry, Kagoshima University Graduate School of Medical and Dental Sciences , Sakuragaoka, Kagoshimashi, Kagoshima , Japan.
authors.forename: Emi
authors.initials: E
authors.lastname: Inada
authors.affiliation: Department of Pediatric Dentistry, Kagoshima University Graduate School of Medical and Dental Sciences , Sakuragaoka, Kagoshimashi, Kagoshima , Japan.
authors.forename: Toshiki
authors.initials: T
authors.lastname: Nomura
authors.affiliation: Department of Pediatric Dentistry, Kagoshima University Graduate School of Medical and Dental Sciences , Sakuragaoka, Kagoshimashi, Kagoshima , Japan.
authors.forename: Eri
authors.initials: E
authors.lastname: Akasaka
authors.affiliation: Department of Pediatric Dentistry, Kagoshima University Graduate School of Medical and Dental Sciences , Sakuragaoka, Kagoshimashi, Kagoshima , Japan.
authors.forename: Youichi
authors.initials: Y
authors.lastname: Yamasaki
authors.affiliation: § Department of Gastroenterological Surgery, Transplant and Surgical Oncology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences , Shikata-cho, Okayama , Japan.
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Noguchi
keywords: ES cell; Feeder; Plasmid; STO cell; Selective drug; iPS cell