Springer US
New York
Springer
http://www.springer-ny.com
11626
10.1007/11626.1543-706X
1071-2690
1543-706X
In Vitro Cellular & Developmental Biology - Animal
In Vitro Cell.Dev.Biol.-Animal
Life Sciences
Cell Biology
Developmental Biology
Stem Cells
Cell Culture
Animal Genetics and Genomics
SC3
52
52
10
2
2
15
2016
2
8
2016
2
8
2016
2
2016
The Society for In Vitro Biology
2016
9968
10.1007/s11626-015-9968-x
15
Generation of cleidocranial dysplasia-specific human induced pluripotent stem cells in completely serum-, feeder-, and integration-free culture
252
264
2015
10
15
2015
10
7
2015
10
15
2015
11
11
Grants-in-Aid for Scientific Research from the Japanese Ministry of Education, Culture, Sports, Science and Technology
24890139
Japan Science and Technology Agency (JP)
AS262Z01770P
The Author(s)
2015
Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
Generation and analysis of CCD-iPSCs
Yamasaki et al.
Sachiko
Yamasaki
Atsuko
Hamada
Eri
Akagi
Hirotaka
Nakatao
Manami
Ohtaka
Ken
Nishimura
Mahito
Nakanishi
Shigeaki
Toratani
Tetsuji
Okamoto
+81-82-257-5665
tetsuok@hiroshima-u.ac.jp
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
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
Laboratory of Gene Regulation, Graduate School of Comprehensive Human Sciences
University of Tsukuba
1-1-1 Tennodai
Tsukuba-shi
Ibaraki
305-8575
Japan
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.
Keywords
iPS cells
Serum-free
Feeder-free
Sendai Virus
Cleidocranial dysplasia
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.