Choice of Feeders is Important When First Establishing iPSCs Derived from Primarily Cultured Human Deciduous Tooth Dental Pulp Cells

Cell Medicine

Yoko Iwase¹  Issei Saitoh¹  Haruaki Hayasaki¹  Tomoya Murakami¹  Miki Soda¹  Kyoko Oka²  Masahiro Sato³  Youichi Yamasaki⁴  Emi Inada⁴  Yuko Matsumoto⁴  Eri Akasaka⁴  Naoko Kubota⁴  Hiroko Hasegawa⁴  Hirofumi Noguchi⁵ 

[1] * Division of Pediatric Dentistry, Graduate School of Medical and Dental Science, Niigata University, Gakkocho-dori, Chuo-ku, Niigata, Japan;§ Section of Pediatric Dentistry Department of Oral Growth and Development Fukuoka Dental College, Sawara-ku, Tamura Fukuoka-shi, Fukuoka, Japan;¶ Section of Gene Expression Regulation, Frontier Science Research Center, Kagoshima University, Sakuragaoka, Kagoshima, Japan;† Department of Pediatric Dentistry, Kagoshima University Graduate School of Medical and Dental Sciences, Sakuragaoka, Kagoshima, Japan;‡ Department of Regenerative Medicine, Graduate School of Medicine, University of the Ryukyus, Nishiharatyoaza, Uehara, Okinawa, Japan

关键词: Deciduous tooth;    Dental pulp;    Feeder cell;    Induced pluripotent stem cells (iPSCs);    Mouse embryonic fibroblasts (MEFs);    STO cells;   

DOI  :  10.3727/215517915X689038

学科分类：生物科学（综合）

来源: Cognizant Communication Corporation

【 摘 要 】

Feeder cells are generally required to maintain embryonic stem cells (ESCs)/induced pluripotent stem cells (iPSCs). Mouse embryonic fibroblasts (MEFs) isolated from fetuses and STO mouse stromal cell line are the most widely used feeder cells. The aim of this study was to determine which cells are suitable for establishing iPSCs from human deciduous tooth dental pulp cells (HDDPCs). Primary cultures of HDDPCs were cotransfected with three plasmids containing human OCT3/4, SOX2/KLF4, or LMYC/LIN28 and pmaxGFP by using a novel electroporation method, and then cultured in an ESC qualified medium for 15 days. Emerging colonies were reseeded onto mitomycin C-treated MEFs or STO cells. The colonies were serially passaged for up to 26 passages. During this period, colony morphology was assessed to determine whether cells exhibited ESC-like morphology and alkaline phosphatase activity to evaluate the state of cellular reprogramming. HDDPCs maintained on MEFs were successfully reprogrammed into iPSCs, whereas those maintained on STO cells were not. Once established, the iPSCs were maintained on STO cells without loss of pluripotency. Our results indicate that MEFs are better feeder cells than STO cells for establishing iPSCs. Feeder choice is a key factor enabling efficient generation of iPSCs.

【 授权许可】

CC BY