We applied scRNA-Seq analysis to elucidate the cellular heterogeneity and explore molecular details better compared with conventional methods. Herein, we used the maxillary process from mouse embryos as a model to understand the development of maxillary-process-derived MSCs. However, application of scRNA-seq in maxillary-process-derived MSCs is still absent. scRNA-seq has been used to study MSCs derived from adipose tissue, bone marrow, endometrium, placenta and dental pulp. scRNA-Seq also provides insights into specific changes in cell lineages, trajectory inference, and the identification of biomarkers. High-throughput single cell RNA sequencing (scRNA-Seq) is powerful for disclosing the complexity and diversity of cells and relationships among genes involved in tissues, and offers an opportunity to explore unbiased gene expression profiling of cells. Nevertheless, accumulating evidence suggests that MSCs are functionally and morphologically heterogeneous in essence. However, the molecular mechanisms of Cd271, as a transmembrane signaling molecule, in the morphogenesis and development of teeth remains largely unknown.Ĭonventionally, MSCs were considered a unified group of fusiform cells. These studies indicate that Cd271 is involved in the physiological processes of tooth development and biomineralization. The spatial–temporal expression of Cd271 was similar to the mineralization factor Runx2 during the early development of tooth germ, which was supported by other studies. Mitsiadis et al indicated that Cd271 surrounded the developing tooth germ within nerve fibers and was highly expressed in the epithelium and mesenchyme in the early stages of odontogenesis within the cells of origin. In addition to these features, emerging studies indicate that Cd271 plays a critical role in initiating tooth development, differentiation and mineralization of odontogenic stem cells. Cd271 is involved in the regulation of morphogenesis and the development of various tissues, including nerves, fat, liver and teeth. Previous studies focused on cellular physiological functions in processes, such as migration, proliferation, differentiation, survival and apoptosis. It is used as a specific cell surface marker for purifying and identifying MSCs. To date, in-depth studies of the odontogenesis and osteogenesis of MSCs are still lacking.Ĭd271 (low-affinity nerve growth factor receptor, p75 neurotrophic receptor) is a member of the tumor necrosis factor receptor superfamily, and is implicated in various biological functions. Ecto-MSCs were regarded as the primitive dental cells in the classical theory of tooth development. Last we verified the findings single cell sequencing in vitro.ĭuring the early stage of embryogenesis, cranial neural crest cells migrate throughout the maxillary and mandibular processes, which are defined as ecto-mesenchymal stem cells (MSCs). We search for marker genes for subgroup annotation by consulting literature and database explore the effect of p75NTR knockout on mesenchymal stem cells (MSCs) gene expression and cell proportion by cell subgrouping, differential gene analysis, enrichment analysis and protein-protein interaction network analysis understand the interaction between MSCs cells and the differentiation trajectory and gene change characteristics of p75NTR knockout MSCs by cell communication analysis and pseudo-time analysis. The gene expression matrix is read by R software, and Seurat is used to control and standardize the data, reduce the dimension and cluster. The FastQC software is used to evaluate the quality of data and CellRanger analyzed the data. Finally, the sequencing data in Fastq format were obtained. After single cell suspension, the cDNA was prepared by loading the single cell suspension into the 10x Genomics Chromium system to be sequenced by NovaSeq6000 sequencing system. P75NTR knockout ( Cd271 -/-) mice (from American Jackson laboratory) were used to collect the maxillofacial process tissue of p75NTR knockout mice, and the wild-type maxillofacial process of the same pregnant mouse wild was used as control.
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