Generation of Mouse STO Feeder Cell Lines that Confer Resistance to Several Types of Selective Drugs

Cell Medicine

Eri Akasaka¹  Youichi Yamasaki¹  Toshiki Nomura¹  Emi Inada¹  Issei Saitoh¹  Hirofumi Noguchi²  Masahiro Sato³  Yoko Iwase⁴ 

[1]* Department of Pediatric Dentistry, Kagoshima University Graduate School of Medical and Dental Sciences, Sakuragaoka, Kagoshimashi, Kagoshima, Japan

[2]§ Department of Gastroenterological Surgery, Transplant and Surgical Oncology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Shikata-cho, Okayama, Japan

[3]† Section of Gene Expression Regulation, Frontier Science Research Center, Kagoshima University, Korimoto, Kagoshima, Kagoshima, Japan

[4]‡ Department of Dental Anesthesia, Kagoshima University Medical and Dental Hospital, Sakuragaoka, Kagoshimashi, Kagoshima, Japan

关键词: Feeder;    STO cell;    Selective drug;    Plasmid;    ES cell;    iPS cell;   

DOI  :  10.3727/215517912X639414

学科分类：生物科学（综合）

来源: Cognizant Communication Corporation

【 摘 要 】

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 phospho-transferase gene (Hyg)], pcBIP [carrying ble and puromycin resistance gene (puro)], or pcBSN [carrying ble and 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.

【 授权许可】

CC BY