title: Insight into the maintenance of odontogenic potential in mouse dental mesenchymal cells based on transcriptomic analysis.
abstract: Background. Mouse dental mesenchymal cells (mDMCs) from tooth germs of cap or later stages are frequently used in the context of developmental biology or whole-tooth regeneration due to their odontogenic potential. In vitro-expanded mDMCs serve as an alternative cell source considering the difficulty in obtaining primary mDMCs; however, cultured mDMCs fail to support tooth development as a result of functional failures of specific genes or pathways. The goal of this study was to identify the genes that maintain the odontogenic potential of mDMCs in culture. Methods. We examined the odontogenic potential of freshly isolated versus cultured mDMCs from the lower first molars of embryonic day 14.5 mice. The transcriptome of mDMCs was detected using RNA sequencing and the data were validated by qRT-PCR. Differential expression analysis and pathway analysis were conducted to identify the genes that contribute to the loss of odontogenic potential. Results. Cultured mDMCs failed to develop into well-structured tooth when they were recombined with dental epithelium. Compared with freshly isolated mDMCs, we found that 1,004 genes were upregulated and 948 were downregulated in cultured mDMCs. The differentially expressed genes were clustered in the biological processes and signaling pathways associated with tooth development. Following in vitro culture, genes encoding a wide array of components of MAPK, TGF-β/BMP, and Wnt pathways were significantly downregulated. Moreover, the activities of Bdnf, Vegfα, Bmp2, and Bmp7 were significantly inhibited in cultured mDMCs. Supplementation of VEGFα, BMP2, and BMP7 restored the expression of a subset of downregulated genes and induced mDMCs to form dentin-like structures in vivo. Conclusions. Vegfα, Bmp2, and Bmp7 play a role in the maintenance of odontogenic potential in mDMCs.
authors.affiliation: Department of Oral and Maxillofacial Surgery, Laboratory of Interdisciplinary Studies, Peking University School and Hospital of Stomatology, Beijing, China.|Department of Orthodontics, Peking University School and Hospital of Stomatology, Beijing, China.
authors.forename: Yunfei
authors.initials: Y
authors.lastname: Zheng
authors.affiliation: Department of Oral and Maxillofacial Surgery, Laboratory of Interdisciplinary Studies, Peking University School and Hospital of Stomatology, Beijing, China.|Department of Orthodontics, Peking University School and Hospital of Stomatology, Beijing, China.|Central Laboratory, Peking University School and Hospital of Stomatology, Beijing, China.
authors.forename: Lingfei
authors.initials: L
authors.lastname: Jia
authors.affiliation: Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China.
authors.forename: Pengfei
authors.initials: P
authors.lastname: Liu
authors.affiliation: Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China.|Experimental Center of Pathogenobiology Immunology, Cytobiology and Genetic, College of Basic Medical Sciences of Jilin University, Jilin, China.
authors.forename: Dandan
authors.initials: D
authors.lastname: Yang
authors.affiliation: Department of Oral and Maxillofacial Surgery, Laboratory of Interdisciplinary Studies, Peking University School and Hospital of Stomatology, Beijing, China.
authors.forename: Waner
authors.initials: W
authors.lastname: Hu
authors.affiliation: Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China.
authors.forename: Shubin
authors.initials: S
authors.lastname: Chen
authors.affiliation: Department of Pediatric Dentistry, Peking University School and Hospital of Stomatology, Beijing, China.
authors.forename: Yuming
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China.
authors.forename: Jinglei
authors.initials: J
authors.lastname: Cai
authors.affiliation: Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China.
authors.forename: Duanqing
authors.initials: D
authors.lastname: Pei
authors.affiliation: Department of Pediatric Dentistry, Peking University School and Hospital of Stomatology, Beijing, China.
authors.forename: Lihong
authors.initials: L
authors.lastname: Ge
authors.affiliation: Department of Oral and Maxillofacial Surgery, Laboratory of Interdisciplinary Studies, Peking University School and Hospital of Stomatology, Beijing, China.|Central Laboratory, Peking University School and Hospital of Stomatology, Beijing, China.
authors.forename: Shicheng
authors.initials: S
authors.lastname: Wei
keywords: Mouse dental mesenchymal cells; Odontogenesis; Odontogenic potential; RNA-Seq; Transcriptome