title: Generation of cleidocranial dysplasia-specific human induced pluripotent stem cells in completely serum-, feeder-, and integration-free culture.
abstract: Human pluripotent stem cells hold great promise for their practical and scientific potentials. To improve understanding of self-renewal and differentiation, we previously reported a defined serum-free medium hESF9 could generate and maintain human induced pluripotent stem cells (iPSCs) in serum- and feeder-free culture conditions using retroviral vectors. To avoid the unpredictable side effects associated with retrovirus integration, we report here the successful generation of hiPSCs from dental pulp cells with a non-integrating replication-defective and persistent Sendai virus (SeVdp) vector expressing four key reprogramming genes. We found that hESF9 medium in combination with fibronectin are effective for generating and maintaining hiPSCs with SeVdp (KOSM). Using this system, pluripotent and self-renewing hiPSCs could be easily and stably generated and propagated. With this system, we successfully generated hiPSCs from cleidocranial dysplasia (CCD) caused by a heterozygous germ-line mutation of runt-related protein2 (RUNX2), which has an important role in the differentiation of osteoblasts and maturation of chondrocytes. This is the first report of the establishment of CCD-specific iPSCs. The cartilage in the teratomas of CCD-iPSCs showed abnormalities. These CCD-iPSCs would be beneficial to clarify the molecular mechanism and for development of medical applications. Moreover, it brings new pathophysiological role of RUNX2 in the differentiation of the human chondrocytes and osteocytes.
authors.affiliation: Department of Molecular Oral Medicine and Maxillofacial Surgery, Applied Life Sciences, Graduate Institute of Biomedical & Health Sciences, Hiroshima University, Kasumi 1-2-3, Minami-ku, Hiroshima, 734-8553, Japan.
authors.forename: Sachiko
authors.initials: S
authors.lastname: Yamasaki
authors.affiliation: Department of Molecular Oral Medicine and Maxillofacial Surgery, Applied Life Sciences, Graduate Institute of Biomedical & Health Sciences, Hiroshima University, Kasumi 1-2-3, Minami-ku, Hiroshima, 734-8553, Japan.
authors.forename: Atsuko
authors.initials: A
authors.lastname: Hamada
authors.affiliation: Department of Molecular Oral Medicine and Maxillofacial Surgery, Applied Life Sciences, Graduate Institute of Biomedical & Health Sciences, Hiroshima University, Kasumi 1-2-3, Minami-ku, Hiroshima, 734-8553, Japan.
authors.forename: Eri
authors.initials: E
authors.lastname: Akagi
authors.affiliation: Department of Molecular Oral Medicine and Maxillofacial Surgery, Applied Life Sciences, Graduate Institute of Biomedical & Health Sciences, Hiroshima University, Kasumi 1-2-3, Minami-ku, Hiroshima, 734-8553, Japan.
authors.forename: Hirotaka
authors.initials: H
authors.lastname: Nakatao
authors.affiliation: Research Center for Stem Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Central 4, Tsukuba, Ibaraki, 305-8562, Japan.
authors.forename: Manami
authors.initials: M
authors.lastname: Ohtaka
authors.affiliation: Laboratory of Gene Regulation, Graduate School of Comprehensive Human Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba-shi, Ibaraki, 305-8575, Japan.
authors.forename: Ken
authors.initials: K
authors.lastname: Nishimura
authors.affiliation: Research Center for Stem Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Central 4, Tsukuba, Ibaraki, 305-8562, Japan.
authors.forename: Mahito
authors.initials: M
authors.lastname: Nakanishi
authors.affiliation: Department of Molecular Oral Medicine and Maxillofacial Surgery, Applied Life Sciences, Graduate Institute of Biomedical & Health Sciences, Hiroshima University, Kasumi 1-2-3, Minami-ku, Hiroshima, 734-8553, Japan.
authors.forename: Shigeaki
authors.initials: S
authors.lastname: Toratani
authors.affiliation: Department of Molecular Oral Medicine and Maxillofacial Surgery, Applied Life Sciences, Graduate Institute of Biomedical & Health Sciences, Hiroshima University, Kasumi 1-2-3, Minami-ku, Hiroshima, 734-8553, Japan. tetsuok@hiroshima-u.ac.jp.
authors.forename: Tetsuji
authors.initials: T
authors.lastname: Okamoto
mesh_terms: D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000235:genetics*; D049109:Cell Proliferation; D002973:Cleidocranial Dysplasia / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D016895:Culture Media, Serum-Free; D003782:Dental Pulp / Q000166:cytology; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects; D029082:Sendai virus / Q000235:genetics
chemical_list: D016895:Culture Media, Serum-Free
keywords: Cleidocranial dysplasia; Feeder-free; Sendai Virus; Serum-free; iPS cells