Project description:To explore the cellular response of stem cell derived from human exfoliated deciduous teeth (SHED) in response to inorganic pyrophosphate (PPi). SHED was extracted from human primary exfoliated teeth. The cellular responses including cell proliferation, apoptosis, migration, transcriptomic expression was investigated afterward PPi treatment at the distinct concentration. Additionally, Alizarin red, oil red O, and alcian blue staining were performed to evaluate the multipotency. Inorganic pyrophosphate (PPi) had no although effect on cell cycle, yet enhanced migratory cell, compared to control. Additionally, PPi abolished the gene expression of osteogenic genes and also calcium deposition. Interestingly, the inhibition of mineralization by PPi was not reversed though Pi3K and MAPK family, including ERK, P38, and JNK signalling pathway. On the contrary, PPi induced the ALPL and COL1A1 gene expression and reduced RANKL mRNA and protein in the condition medium, resulting in a decrease of osteoclast formation. The transcriptomic profiles illustrated PPi modulated interferon ⍺/ , TGF-β1, NOTCH signalling pathway, and metabolism of lipid. In summary, our study revealed that PPi enhanced migratory ability, however inhibited osteo/odontogenic, adipogenic and chondrogenic differentiation. This study implicated that PPi can modulate cellular responses of SHED.
Project description:bFGF regulated stemness maintenance and cell differentiation in stem cells isolated from human exfoliated deciduous teeth (SHEDs). In addition, WNT3A is also shown to regulate osteogenic differentiation in mesenchymal stem cells. Here, transcriptome analysis of bFGF and WNT3A treated SHEDs was evaluated using a high throughput RNA sequencing technique. Results demonstrated that bFGF regulated genes related to cell cycle control and cell proliferation in SHEDs.
Project description:We employed microarray profiling to determine the gene(s) that control human exfoliated deciduous teeth (SHED) differentiation into epithelial-like cells. We introduced epithelial microenvironment using keratinocyte serum free medium (KSFM) and co-culture with human epithelial keratinocyte (HEK001) to promote epithelial-like differentiation.
Project description:Stem cells from human exfoliated deciduous teeth (SHED) can be used as a potential clinical material. But the use of xenogeneic ingredients, especially fetal bovine serum, will increase the risk of zoonotic disease transmission during the use of animal-derived products. Human-platelet-lysate (hPL) was used in human cell expansion with reliability in clinical applications. This study aimed to explore the feasibility and effect of HPL in the cultivation of SHED. We use proteomics to interpret the impact of HPL on the protein profile of SHED. Propagation, differentiation, qPCR and immunofluorescence were used to detect the effect of HPL on SHED.
Project description:Human deciduous and permanent teeth exhibit different developmental processes, morphologies, histological characteristics and life cycles. In addition their pulp tissues react differently to external stimuli, such as the pulp sensitivity test, dental trauma and pulp therapy materials. These differences are attributable to their genetic backgrounds. Therefore the purpose of this study is to compare the differences of dental pulp in deciduous and permanent teeth. Pulp samples were obtained from permanent premolars (n=6, aged 11-14 years) and deciduous teeth (n=6, aged 11-14 years). Comparative cDNA microarrary analysis revealed several differences in gene expression between the deciduous and permanent pulp tissues. Each GSM record represents a pulp sample pooled from two teeth samples.
Project description:Deciduous and permanent human teeth represent a model system to study ageing of mesenchymal populations. Aging is tightly connected to self-renewal and proliferation and thus, mapping potential molecular differences in these characteristics between populations constitutes an important task. Specifically designed microarray panels were used. We have detected a number of molecules that were differentially expressed in dental pulp mesenchyme from deciduous and permanent teeth extracted from young children and adults, respectively. Among the differentially regulated genes HMGA2, a stem cell-associated marker, stood out as a remarkable example with a robust expression in deciduous pulp cells. In addition to this, we discovered that several proliferation-related genes, including CDC2A and CDK4, were up-regulated in deciduous pulp cells, while matrix genes COL1A1, fibronectin and several signaling molecules, such as VEGF, FGFr-1 and IGFr-1 were up-regulated in the pulp cells from permanent teeth. Taken together, our data suggest that deciduous pulp cells are more robust in self- renewal and proliferation, whereas adult dental pulp cells are more capable of signaling and matrix synthesis.
Project description:Human deciduous and permanent teeth exhibit different developmental processes, morphologies, histological characteristics and life cycles. In addition their pulp tissues react differently to external stimuli, such as the pulp sensitivity test, dental trauma and pulp therapy materials. These differences are attributable to their genetic backgrounds. Therefore the purpose of this study is to compare the differences of dental pulp in deciduous and permanent teeth.
Project description:Deciduous and permanent human teeth represent a model system to study ageing of mesenchymal populations. Aging is tightly connected to self-renewal and proliferation and thus, mapping potential molecular differences in these characteristics between populations constitutes an important task. Specifically designed microarray panels were used. We have detected a number of molecules that were differentially expressed in dental pulp mesenchyme from deciduous and permanent teeth extracted from young children and adults, respectively. Among the differentially regulated genes HMGA2, a stem cell-associated marker, stood out as a remarkable example with a robust expression in deciduous pulp cells. In addition to this, we discovered that several proliferation-related genes, including CDC2A and CDK4, were up-regulated in deciduous pulp cells, while matrix genes COL1A1, fibronectin and several signaling molecules, such as VEGF, FGFr-1 and IGFr-1 were up-regulated in the pulp cells from permanent teeth. Taken together, our data suggest that deciduous pulp cells are more robust in self- renewal and proliferation, whereas adult dental pulp cells are more capable of signaling and matrix synthesis. The deciduous teeth were collected from 3-12-year-old children (N=8), whereas the permanent teeth were collected from adults, 19-52 years of age (N=8). The tooth was mechanically separated into two halves to remove all soft tissue in the pulp. The pulp tissue was digested and the dissociated cells were passed through cell strainer and plated with culture medium. The cells were then passaged at 90% confluency to a 10 cm dish for expansion. RNA was extracted from passage one cells with the Qiagen RNeasy Mini Kit. Microarray analysis was performed using PIQORTM Stem Cell Microarray chip (Miltenyi Biotech), which is comprised of 942 relevant genes for stem/progenitor cells and key genes for cell differentiation, to identify gene sets that are differentially expressed between cells in the deciduous and adult teeth. A total of 1 μg RNA for each sample was used for amplification and further analysis with the PIQORTM stem cell microarray chip (Miltenyi Biotec), followed by detection with a laser scanner (Agilent Technologies). In total, 8 comparisons were performed with dataset consisting of two microarrays, each containing a total of 11 slides with signal intensities within the R/Bioconductor statistical framework. The Linear Model for Microarray Data package (Limma) was used to pre-process the raw data, perform quality controls and estimate statistical significance. Expression intensities were background-corrected using a convolution of normal and exponential distributions with an offset of 50, which were then normalized within every slide using loess normalization. We analyzed the red and green channels separately using the mixed model method and different groups were quantile-normalized separately. The correlation was computed between the two channels for the same spot. Differentially expressed genes were identified by computing the contrasts, fitting to a linear model, performing the hypothesis tests, and correcting for multiple testing by performing the false discovery rate using the Benjamini-Hochberg correction.
Project description:Background: Mesenchymal stem cells (MSCs) have been widely used in the treatment of various inflammatory diseases. The inadequate understanding of MSCs and its heterogeneity can impact the immune environment maybe the cause of the good outcomes of MSCs-based therapy cannot be always achieved. Recently, stem cells from human exfoliated deciduous teeth (SHED) showed a great potential in inflammatory and autoimmune diseases due to its unique immunomodulation properties. However, the cellular heterogeneity and the corresponding immunomodulation functions of SHED remains unclear. Methods: In this study, we applied single-cell RNA sequencing (scRNA-seq) to analyze and performed bioinformatic analysis to clarify the variations immunomodulation functions of SHED subpopulations in different differentiation state. After the integration of public available databases, we analyzed the transcriptome and compared the difference of immunomodulation functions of MSCs from different tissues. Results: SHED in low differentiation state (S7) exhibited the powerful ability to recruit multiple types of immune cells, whereas SHED in relatively high differentiation state (S1) may hold strong ability to secret many factors with paracrine signaling capacity. Compared with human bone marrow derived mesenchymal stem cells (hBMSCs) or human umbilical cord-derived mesenchymal stem cells (hUCMSCs), SHED is stronger in immunomodulatory property, especially in recruitment of Th17, Th22 and Treg cells. Conclusion: SHED was a heterogeneous MSCs population with different differentiation states and immunomodulatory functions. SHED may have some advantages in treatment of inflammatory and autoimmune diseases. Our works provides some theoretical foundation for the SHED-based therapy in clinical applications.