title: Choice of Feeders Is Important When First Establishing iPSCs Derived From Primarily Cultured Human Deciduous Tooth Dental Pulp Cells.
abstract: 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.
authors: Division of Pediatric Dentistry, Graduate School of Medical and Dental Science, Niigata University , Gakkocho-dori, Chuo-ku, Niigata , Japan.
authors: Issei
authors: I
authors: Saitoh
authors: † Department of Pediatric Dentistry, Kagoshima University Graduate School of Medical and Dental Sciences , Sakuragaoka, Kagoshima , Japan.
authors: Emi
authors: E
authors: Inada
authors: Division of Pediatric Dentistry, Graduate School of Medical and Dental Science, Niigata University , Gakkocho-dori, Chuo-ku, Niigata , Japan.
authors: Yoko
authors: Y
authors: Iwase
authors: ‡ Department of Regenerative Medicine, Graduate School of Medicine, University of the Ryukyus , Nishiharatyoaza, Uehara, Okinawa , Japan.
authors: Hirofumi
authors: H
authors: Noguchi
authors: Division of Pediatric Dentistry, Graduate School of Medical and Dental Science, Niigata University , Gakkocho-dori, Chuo-ku, Niigata , Japan.
authors: Tomoya
authors: T
authors: Murakami
authors: Division of Pediatric Dentistry, Graduate School of Medical and Dental Science, Niigata University , Gakkocho-dori, Chuo-ku, Niigata , Japan.
authors: Miki
authors: M
authors: Soda
authors: † Department of Pediatric Dentistry, Kagoshima University Graduate School of Medical and Dental Sciences , Sakuragaoka, Kagoshima , Japan.
authors: Naoko
authors: N
authors: Kubota
authors: † Department of Pediatric Dentistry, Kagoshima University Graduate School of Medical and Dental Sciences , Sakuragaoka, Kagoshima , Japan.
authors: Hiroko
authors: H
authors: Hasegawa
authors: † Department of Pediatric Dentistry, Kagoshima University Graduate School of Medical and Dental Sciences , Sakuragaoka, Kagoshima , Japan.
authors: Eri
authors: E
authors: Akasaka
authors: † Department of Pediatric Dentistry, Kagoshima University Graduate School of Medical and Dental Sciences , Sakuragaoka, Kagoshima , Japan.
authors: Yuko
authors: Y
authors: Matsumoto
authors: § Section of Pediatric Dentistry Department of Oral Growth and Development Fukuoka Dental College , Sawara-ku, Tamura Fukuoka-shi, Fukuoka , Japan.
authors: Kyoko
authors: K
authors: Oka
authors: † Department of Pediatric Dentistry, Kagoshima University Graduate School of Medical and Dental Sciences , Sakuragaoka, Kagoshima , Japan.
authors: Youichi
authors: Y
authors: Yamasaki
authors: Division of Pediatric Dentistry, Graduate School of Medical and Dental Science, Niigata University , Gakkocho-dori, Chuo-ku, Niigata , Japan.
authors: Haruaki
authors: H
authors: Hayasaki
authors: ¶ Section of Gene Expression Regulation, Frontier Science Research Center, Kagoshima University , Sakuragaoka, Kagoshima , Japan.
authors: Masahiro
authors: M
authors: Sato
keywords: Deciduous tooth; Dental pulp; Feeder cell; Induced pluripotent stem cells (iPSCs); Mouse embryonic fibroblasts (MEFs); STO cells