Optimization of culture conditions for the efficient differentiation of mouse-induced pluripotent stem cells into dental
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Optimization of culture conditions for the efficient differentiation of mouse-induced pluripotent stem cells into dental epithelial-like cells Azusa Onishi 1,2 & Aimi Naim Abdullah 1,2 & Kotaro Tanimoto 2 & Koichi Kato 1 Received: 8 May 2020 / Accepted: 2 September 2020 / Editor: Tetsuji Okamoto # The Society for In Vitro Biology 2020
Abstract The establishment of a method to derive dental epithelial cells seems to be an important challenge toward realizing the whole tooth regeneration. In order to obtain a source of dental epithelial-like cells, a new methodology has been previously developed by our research group. In the method, induced pluripotent stem cells are cultured in suspension in the presence of neurotrophin-4 to form embryoid bodies followed by further adherent culture of the embryoid bodies in DMEM basal nutrient medium. The present study was directed to improve the efficiency of dental epithelial-like cell production, by focusing on the optimization of initial cell number for the formation of embryoid bodies and the addition of epidermal growth factor as well as its timing. Our results demonstrated that an initial cell number of 1000 cells/drop gives the highest efficiency of dental epithelial-like cell production. It appears that, under this condition, medium deterioration is moderated, and that cell-cell interactions are optimized within embryoid bodies. On the other hand, epidermal growth factor serves to increase the abundance of dental epithelial-like cells when added to the medium together with neurotrophin-4 during embryoid body formation. The promotive effect of epidermal growth factor may involve the transactivation of TrkB, mediated by the effectors of epidermal growth factor receptor signaling. Keywords Induced pluripotent stem cell . Tooth regeneration . Epithelial cell . Ameloblast . Epidermal growth factor
Introduction Dental epithelial cells are found at the early stage of tooth development and play a crucial role in tooth formation through the reciprocal interaction with mesenchymal cells. Dental epithelial cells are lost during tooth eruption and are not available in adult tissues (Colin et al. 1998; Mitsiadis and Pappagerakis 2001; Bluteau et al. 2008; Lacruz et al. 2017). Recently, it was shown that dental epithelial cells obtained from fetal tissues have potential to duplicate tooth-like structure when combined
* Koichi Kato [email protected] 1
Department of Biomaterials, Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553, Japan
2
Department of Orthodontics and Craniofacial Developmental Biology, Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553, Japan
with immature mesenchymal cells (Duailibi et al. 2004; Sonoyama et al. 2006; Honda et al. 2007; Ikeda et al. 2009; Oshima et al. 2011). This finding opened a new venue for developing a strategy to regenerate a whole tooth in a patient suffering from missing teeth. However, as just describe
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