A single-cell transcriptomic atlas of human stem cells from apical papilla during the committed differentiation.

IF 5.4 1区 医学 Q1 DENTISTRY, ORAL SURGERY & MEDICINE International endodontic journal Pub Date : 2024-11-12 DOI:10.1111/iej.14170
Yingying Weng, Ya Xiao, Yijia Shi, Na Li, Jing Wang, Ming Yan, Jinhua Yu, Zehan Li
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Abstract

Aim: Human stem cells derived from the apical papilla (SCAPs) are recognized for their multilineage differentiation potential and their capacity for functional tooth root regeneration. However, the molecular mechanisms underlying odonto/osteogenic differentiation remain largely unexplored. In this study, we utilized single-cell RNA sequencing (scRNA-seq) to conduct an in-depth analysis of the transcriptional changes associated with chemically induced osteogenesis in SCAPs.

Methodology: scRNA-seq identified SCAPs as distinct subpopulations. Differentially expressed genes (DEGs) and Gene Ontology (GO) analyses were conducted to evaluate the potential function of each cluster. Pseudotime trajectory analysis was employed to elucidate the potential differentiation processes of the identified SCAP populations. To investigate the osteo/odontogenic potential of Deiodinase Iodothyronine Type 2 (DIO2) on SCAPs, we performed alkaline phosphatase staining, western blot analysis, Alizarin Red S staining and immunofluorescence staining. Additionally, SCAP components were transplanted into mouse calvarial defects to evaluate osteogenesis in vivo.

Results: The analysis of cell clusters derived from our scRNA-seq data revealed a significant shift in cellular composition when cells were cultured in a mineralization induction medium compared to those cultured in a complete medium. Both groups exhibited heterogeneity, with some cells intrinsically predisposed to osteogenesis and others appearing to be primed for proliferative functions. Notably, we identified a subpopulation characterized by high expression of DIO2, which exhibited pronounced osteogenic activity during differentiation.

Conclusions: Our study is the first to reveal a shift in the cellular composition of SCAPs when cultured in a mineralization induction medium compared to a complete medium. Following both in vitro and in vivo validation, the DIO2+ subpopulation exhibited the highest transcriptional similarity to osteogenic function, suggesting its potential utility in tissue regeneration applications.

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来自顶端乳头的人类干细胞在定向分化过程中的单细胞转录组图谱。
目的:源自牙根尖乳头的人类干细胞(SCAPs)被认为具有多线分化潜力和功能性牙根再生能力。然而,牙髓/牙根分化的分子机制在很大程度上仍未得到探索。在这项研究中,我们利用单细胞 RNA 测序(scRNA-seq)对 SCAPs 中与化学诱导成骨相关的转录变化进行了深入分析。进行了差异表达基因(DEGs)和基因本体(GO)分析,以评估每个群的潜在功能。采用伪时间轨迹分析来阐明已确定的 SCAP 群体的潜在分化过程。为了研究碘甲腺原氨酸酶 2 型(DIO2)在 SCAP 上的成骨/成牙潜力,我们进行了碱性磷酸酶染色、Western 印迹分析、茜素红 S 染色和免疫荧光染色。此外,我们还将 SCAP 成分移植到小鼠腓骨缺损处,以评估体内成骨情况:从 scRNA-seq 数据中得出的细胞群分析表明,在矿化诱导培养基中培养的细胞与在完全培养基中培养的细胞相比,细胞组成发生了显著变化。两组细胞都表现出异质性,一些细胞天生倾向于成骨,而另一些细胞似乎具有增殖功能。值得注意的是,我们发现了一个以高表达 DIO2 为特征的亚群,该亚群在分化过程中表现出明显的成骨活性:我们的研究首次揭示了与完全培养基相比,在矿化诱导培养基中培养 SCAPs 时细胞组成的变化。经过体外和体内验证,DIO2+亚群表现出与成骨功能最高的转录相似性,这表明它在组织再生应用中具有潜在的实用性。
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来源期刊
International endodontic journal
International endodontic journal 医学-牙科与口腔外科
CiteScore
10.20
自引率
28.00%
发文量
195
审稿时长
4-8 weeks
期刊介绍: The International Endodontic Journal is published monthly and strives to publish original articles of the highest quality to disseminate scientific and clinical knowledge; all manuscripts are subjected to peer review. Original scientific articles are published in the areas of biomedical science, applied materials science, bioengineering, epidemiology and social science relevant to endodontic disease and its management, and to the restoration of root-treated teeth. In addition, review articles, reports of clinical cases, book reviews, summaries and abstracts of scientific meetings and news items are accepted. The International Endodontic Journal is essential reading for general dental practitioners, specialist endodontists, research, scientists and dental teachers.
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