title: Derivation of iPSCs after culture of human dental pulp cells under defined conditions.
abstract: Human dental pulp cells (hDPCs) are a promising resource for regenerative medicine and tissue engineering and can be used for derivation of induced pluripotent stem cells (iPSCs). However, current protocols use reagents of animal origin (mainly fetal bovine serum, FBS) that carry the potential risk of infectious diseases and unwanted immunogenicity. Here, we report a chemically defined protocol to isolate and maintain the growth and differentiation potential of hDPCs. hDPCs cultured under these conditions showed significantly less primary colony formation than those with FBS. Cell culture under stringently defined conditions revealed a donor-dependent growth capacity; however, once established, the differentiation capabilities of the hDPCs were comparable to those observed with FBS. DNA array analyses indicated that the culture conditions robustly altered hDPC gene expression patterns but, more importantly, had little effect on neither pluripotent gene expression nor the efficiency of iPSC induction. The chemically defined culture conditions described herein are not perfect serum replacements, but can be used for the safe establishment of iPSCs and will find utility in applications for cell-based regenerative medicine.
authors.affiliation: Department of Oral and Maxillofacial Science, Gifu University Graduate School of Medicine, Gifu, Japan.
authors.forename: Tomoko
authors.initials: T
authors.lastname: Takeda-Kawaguchi
authors.affiliation: Department of Oral and Maxillofacial Science, Gifu University Graduate School of Medicine, Gifu, Japan.
authors.forename: Ken
authors.initials: K
authors.lastname: Sugiyama
authors.affiliation: Department of Tissue and Organ Development, Gifu University Graduate School of Medicine, Gifu, Japan.
authors.forename: Shunji
authors.initials: S
authors.lastname: Chikusa
authors.affiliation: Department of Oral and Maxillofacial Science, Gifu University Graduate School of Medicine, Gifu, Japan.
authors.forename: Kazuki
authors.initials: K
authors.lastname: Iida
authors.affiliation: Department of Tissue and Organ Development, Gifu University Graduate School of Medicine, Gifu, Japan.
authors.forename: Hitomi
authors.initials: H
authors.lastname: Aoki
authors.affiliation: Department of Oral and Maxillofacial Science, Gifu University Graduate School of Medicine, Gifu, Japan.
authors.forename: Naritaka
authors.initials: N
authors.lastname: Tamaoki
authors.affiliation: Department of Oral and Maxillofacial Science, Gifu University Graduate School of Medicine, Gifu, Japan.
authors.forename: Daijiro
authors.initials: D
authors.lastname: Hatakeyama
authors.affiliation: Department of Tissue and Organ Development, Gifu University Graduate School of Medicine, Gifu, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Kunisada
authors.affiliation: Department of Oral and Maxillofacial Science, Gifu University Graduate School of Medicine, Gifu, Japan.
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Shibata
authors.affiliation: Department of Ophthalmology, Keio University School of Medicine, Tokyo, Japan.
authors.forename: Noemi
authors.initials: N
authors.lastname: Fusaki
authors.affiliation: Department of Tissue and Organ Development, Gifu University Graduate School of Medicine, Gifu, Japan.
authors.forename: Ken-Ichi
authors.initials: K
authors.lastname: Tezuka
mesh_terms: D000293:Adolescent; D002454:Cell Differentiation; D002478:Cells, Cultured; D003782:Dental Pulp / Q000166:cytology*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D061251:Primary Cell Culture / Q000379:methods