Single-cell RNA-seq of in vitro expanded cells from cranial neural crest reveals a rare odontogenic sub-population

Yifan Zhao; Shubin Chen; Xiaobo Liu; Xiaoming Chen; Dandan Yang; Jiashu Zhang; Di Wu; Yanmei Zhang; Si Xie; Xiaomei Li; Zhiyuan Wang; Bo Feng; Dajiang Qin; Duanqing Pei; Yaofeng Wang; Jinglei Cai

doi:10.1111/cpr.13598

Cell Proliferation ›› 2024, Vol. 57 ›› Issue (6) :e13598 DOI: 10.1111/cpr.13598

ORIGINAL ARTICLE

Single-cell RNA-seq of in vitro expanded cells from cranial neural crest reveals a rare odontogenic sub-population

Yifan Zhao ¹^,²^,³^,⁴
, Shubin Chen ¹^,³^,⁴
, Xiaobo Liu ⁵
, Xiaoming Chen ⁴^,⁶
, Dandan Yang ⁷
, Jiashu Zhang ⁸
, Di Wu ³^,⁴
, Yanmei Zhang ⁴
, Si Xie ¹^,²
, Xiaomei Li ¹
, Zhiyuan Wang ⁸
, Bo Feng ⁹
, Dajiang Qin ¹^,²^,³
, Duanqing Pei ²^,¹⁰
, Yaofeng Wang ¹^,²^,^†
, Jinglei Cai ¹^,²^,⁴^,¹¹^,¹²^,^†

Author information +

¹. Innovation Centre for Advanced Interdisciplinary Medicine, The Fifth Affiliated Hospital of Guangzhou Medical University, Guangzhou, China

². Centre for Regenerative Medicine and Health, Hong Kong Institute of Science & Innovation, Chinese Academy of Sciences, Hong Kong SAR, China

³. Bioland Laboratory, Guangzhou Regenerative Medicine and Health Guangdong Laboratory, Guangzhou, China

⁴. CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China

⁵. Laboratory of Cancer Precision Medicine, The First Hospital of Jilin University, Changchun, China

⁶. Guangdong Provincial People's Hospital Ganzhou Hospital, Ganzhou Municipal Hospital, Ganzhou, China

⁷. Experimental Center of Pathogenobiology Immunology, Cytobiology and Genetics, Basic Medical College, Jilin University, Changchun, China

⁸. Innovation Centre for Translational Medicine, The Fifth Affiliated Hospital of Guangzhou Medical University, Guangzhou, China

⁹. School of Biomedical Sciences, The Chinese University of Hong Kong, Hong Kong SAR, China

¹⁰. Laboratory of Cell Fate Control, School of Life Sciences, Westlake University, Hangzhou, China

¹¹. Guangzhou Key Laboratory of Enhanced Recovery after Abdominal Surgery, The Fifth Affiliated Hospital of Guangzhou Medical University, Guangzhou, China

¹². Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing, China

yaofeng.wang@hkisi-cas.org.hk

caijinglei@gzhmu.edu.cn

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Abstract

Ecto-mesenchymal cells of mammalian tooth germ develops from cranial neural crest cells. These cells are recognised as a promising source for tooth development and regeneration. Despite the high heterogeneity of the neural crest, the cellular landscape of in vitro cultured cranial neural crest cells (CNCCs) for odontogenesis remains unclear. In this study, we used large-scale single-cell RNA sequencing to analyse the cellular landscape of in vitro cultured mouse CNCCs for odontogenesis. We revealed distinct cell trajectories from primary cells to passage 5 and identified a rare Alx3+/Barx1+ sub-population in primary CNCCs that differentiated into two odontogenic clusters characterised by the up-regulation of Pax9/Bmp3 and Lhx6/Dmp1. We successfully induced whole tooth-like structures containing enamel, dentin, and pulp under the mouse renal capsule using in vitro cultured cells from both cranial and trunk neural crests with induction rates of 26.7% and 22.1%, respectively. Importantly, we confirmed only cells sorted from odontogenic path can induce tooth-like structures. Cell cycle and DNA replication genes were concomitantly upregulated in the cultured NCCs of the tooth induction groups. Our data provide valuable insights into the cell heterogeneity of in vitro cultured CNCCs and their potential as a source for tooth regeneration.

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Yifan Zhao, Shubin Chen, Xiaobo Liu, Xiaoming Chen, Dandan Yang, Jiashu Zhang, Di Wu, Yanmei Zhang, Si Xie, Xiaomei Li, Zhiyuan Wang, Bo Feng, Dajiang Qin, Duanqing Pei, Yaofeng Wang, Jinglei Cai. Single-cell RNA-seq of in vitro expanded cells from cranial neural crest reveals a rare odontogenic sub-population. Cell Proliferation, 2024, 57(6): e13598 DOI:10.1111/cpr.13598

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