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Open AccessArticle

In Vitro Characterization of Dental Pulp Stem Cells Cultured in Two Microsphere-Forming Culture Plates

1
Department of Conservative Dentistry, Graduate School, Kyung Hee University, Seoul 02447, Korea
2
Department of Pediatric Dentistry, School of Dentistry, Kyung Hee University, Seoul 02447, Korea
3
Department of Conservative Dentistry, School of Dentistry, Dental Science Research Institute, Chonnam National University, Gwang-ju 61186, Korea
4
Department of Conservative Dentistry and Dental Research Institute, School of Dentistry, Seoul National University, Seoul 03080, Korea
5
Department of Conservative Dentistry, School of Dentistry, Kyung Hee University, Seoul 02447, Korea
*
Author to whom correspondence should be addressed.
J. Clin. Med. 2020, 9(1), 242; https://doi.org/10.3390/jcm9010242 (registering DOI)
Received: 26 December 2019 / Revised: 10 January 2020 / Accepted: 13 January 2020 / Published: 16 January 2020
(This article belongs to the Special Issue Innovations in Endodontic Dentistry)
Various three-dimensional (3D) culture methods have been introduced to overcome the limitations of in vitro culture and mimic in vivo conditions. This study aimed to evaluate two microsphere-forming culture methods and a monolayer culture method. We evaluated cell morphology, viability, osteo-, adipo-, and chondrogenic differentiation potential of dental pulp stem cells (DPSCs) cultured in 3D culture plates: ultra-low attachment (ULA) and U-bottomed StemFit 3D (SF) plates, and a two-dimensional (2D) monolayer plate. RNA sequencing (RNA-seq) revealed differentially expressed gene (DEG) profiles of the DPSCs. In contrast to an increasing pattern in the 2D group, cell viability in 3D groups (ULA and SF) showed a decreasing pattern; however, high multilineage differentiation was observed in both the 3D groups. RNA-seq showed significantly overexpressed gene ontology categories including angiogenesis, cell migration, differentiation, and proliferation in the 3D groups. Hierarchical clustering analysis revealed a similar DEG regulation pattern between the 3D groups; however, a comparatively different DEG was observed between the 2D and 3D groups. Taken together, this study shows that DPSCs cultured in microsphere-forming plates present superior multilineage differentiation capacities and demonstrate higher DEG expression in regeneration-related gene categories compared to that in DPSCs cultured in a conventional monolayer plate. View Full-Text
Keywords: culture method; dental pulp; dental pulp stem cell; microsphere; multilineage differentiation; pulp regeneration; RNA sequencing culture method; dental pulp; dental pulp stem cell; microsphere; multilineage differentiation; pulp regeneration; RNA sequencing
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Bu, N.-U.; Lee, H.-S.; Lee, B.-N.; Hwang, Y.-C.; Kim, S.-Y.; Chang, S.W.; Choi, K.-K.; Kim, D.-S.; Jang, J.-H. In Vitro Characterization of Dental Pulp Stem Cells Cultured in Two Microsphere-Forming Culture Plates. J. Clin. Med. 2020, 9, 242.

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