Generation of cleidocranial dysplasia-specific human induced pluripotent stem cells in completely serum-, feeder-, and integration-free culture

Citation metadata

Publisher: Springer
Document Type: Report
Length: 367 words

Document controls

Main content

Abstract :

Byline: Sachiko Yamasaki (1), Atsuko Hamada (1), Eri Akagi (1), Hirotaka Nakatao (1), Manami Ohtaka (2), Ken Nishimura (3), Mahito Nakanishi (2), Shigeaki Toratani (1), Tetsuji Okamoto (1) Keywords: iPS cells; Serum-free; Feeder-free; Sendai Virus; Cleidocranial dysplasia 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. Author Affiliation: (1) 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 (2) 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 (3) Laboratory of Gene Regulation, Graduate School of Comprehensive Human Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba-shi, Ibaraki, 305-8575, Japan Article History: Registration Date: 15/10/2015 Received Date: 07/10/2015 Accepted Date: 15/10/2015 Online Date: 11/11/2015 Article note: Editor: J. Denry Sato Sachiko Yamasaki and Atsuko Hamada contributed equally to this work. Electronic supplementary material The online version of this article (doi: 10.1007/s11626-015-9968-x) contains supplementary material, which is available to authorized users.

Source Citation

Source Citation   

Gale Document Number: GALE|A442791036