human dpscs  (Celprogen Inc)

Journal: Frontiers in Genetics

Article Title: Gene expression profiles identify key factors in inflammatory odontoblastic dental pulp stem cell differentiation via TNFα/C5L2

doi: 10.3389/fgene.2025.1592599

Figure Lengend Snippet: Analysis of differentially expressed genes (DEGs) in RNA-Seq. data. in TNFα-treated odontoblast-like differentiating DPSCs. (A) Heatmap of the differentially expressed genes between untreated or control cells and TNFα hDPSCs in dentinogenic media. Red and yellow stripes in the figure represent high-expression genes, while blue stripes represent low-expression genes. Particular genes and transcription factors of interest that are involved in odontoblastic differentiation are red circled from the clusters and shown in the following figure in the graphs. (B) Volcano map of differentially expressed genes (DEGs) between control cells and TNFα hDPSCs in dentinogenic media. The x-axis is the log2 scale of the fold change of gene expression in hDPSCs (log2 (fold change)). Negative values indicate downregulation; positive values indicate upregulation. The y-axis is the minus log10 scale of the adjusted p values (–log10), indicating the significant expression difference level. The blue dots represent significantly upregulated genes with at least twofold change, while the red dots represent significantly downregulated genes with at least twofold change. Key genes have been mentioned along the volcano dots. (C) Based on their functions, significantly enriched Gene Ontology (GO) terms were found among control and TNFα-treated hDPSCs. The top GO terms in the enrichment analysis are biological process, cellular component, and molecular function (MF) terms in the enrichment analysis.

Article Snippet: Human DPSCs were purchased from Lonza, Pharma & Biotech (Cat. # PT-5025).

Techniques: RNA Sequencing, Control, Expressing, Gene Expression

Journal: Frontiers in Genetics

Article Title: Gene expression profiles identify key factors in inflammatory odontoblastic dental pulp stem cell differentiation via TNFα/C5L2

doi: 10.3389/fgene.2025.1592599

Figure Lengend Snippet: Signaling pathways affected by TNFα in odontoblast-like differentiation of hDPSCs in dentinogenic media. hDPSCs were cultured and treated with or without TNFα for 7 days (twice a week with 3-day intervals). Cell lysates were collected, and RNA was prepared using the RNeasy mini kit (Qiagen), and next-generation RNA sequencing was done using the poly-A-RNA sequencing technique. (A) Histogram showing upregulated and activated transcription factors (blue) and repressed or downregulated transcription factors (orange). Notably, TCF ( ; ; ; ; ), especially TCF12, is highly upregulated in TNFα-stimulated odontoblasts like differentiated DPSCs. (B) Histogram showing upregulated and activated upregulated genes (blue) and repressed or downregulated genes (orange). Notably, key genes involved in dentinogenesis are significantly upregulated in TNFα-stimulated odontoblast-like differentiated DPSCs.

Article Snippet: Human DPSCs were purchased from Lonza, Pharma & Biotech (Cat. # PT-5025).

Techniques: Protein-Protein interactions, Cell Culture, RNA Sequencing

Journal: Frontiers in Genetics

Article Title: Gene expression profiles identify key factors in inflammatory odontoblastic dental pulp stem cell differentiation via TNFα/C5L2

doi: 10.3389/fgene.2025.1592599

Figure Lengend Snippet: Analysis of differentially expressed genes (DEGs) in RNA-Seq. data in siC5L2-treated odontoblast-like differentiating DPSCs. (A) Heatmap of the differentially expressed genes between untreated or control cells and siC5L2-treated hDPSCs in dentinogenic media. Red and yellow stripes in the figure represent high-expression genes, while blue stripes represent low-expression genes. Particular genes and transcription factors of interest that are involved in odontoblastic differentiation are red circled from the clusters and shown in the following figure in the graphs. (B) Volcano map of differentially expressed genes (DEGs) between control cells and siC5L2 hDPSCs in dentinogenic media. The x-axis is the log2 scale of the fold change of gene expression in hDPSCs (log2 (fold change)). Negative values indicate downregulation; positive values indicate upregulation. The y-axis is the minus log10 scale of the adjusted p values (–log10), indicating the significant expression difference level. The blue dots represent significantly upregulated genes with at least twofold change, while the red dots represent significantly downregulated genes with at least twofold change. Key genes have been mentioned along the volcano dots. (C) Their functions are significantly enriched in Gene Ontology (GO) terms among control and siC5L2-treated hDPSCs.

Article Snippet: Human DPSCs were purchased from Lonza, Pharma & Biotech (Cat. # PT-5025).

Techniques: RNA Sequencing, Control, Expressing, Gene Expression

Journal: Frontiers in Genetics

Article Title: Gene expression profiles identify key factors in inflammatory odontoblastic dental pulp stem cell differentiation via TNFα/C5L2

doi: 10.3389/fgene.2025.1592599

Figure Lengend Snippet: The top GO terms in the enrichment analysis among biological process, cellular component, and molecular function (MF) terms in the enrichment analysis in siC5L2-treated odontoblast-like differentiating DPSCs. Significantly enriched GO terms can be found in TNFα-treated hDPSCs in dentinogenic media based on their functions compared with the control. (A,B) Go-term and Reactome enrichment pathway analysis of up- and downregulated DEGs. The dot plot shows the top enriched Reactome pathways. The dot size is based on the gene count enriched in the pathway, and the dot color shows the significance of pathway enrichment. (C) Gene set enrichment analysis was performed on DEGs among control and siC5L2-treated cells in dentinogenic media, and various upregulated and downregulated genes were found to be against bacterial response. Enrichment plot and Random ES distribution show that the defense response against bacteria has been enhanced after C5L2 silencing. (D) Sashimi plots for quantitative visualization of RNA sequencing read alignments. The arched lines indicate splice junctions; the number of reads using that splice junction is indicated on the arched line. Data were examined on sashimi plots, revealing variants and genomic mutations on chr16, chr19, and chr11 in siC5L2-treated cells in dentinogenic media against the control. Red sashimi plots show variants in the siC5L2-treated group, and orange shows in the control. While lower black annotations are Read alignments of alternative isoforms and genomic regions of interest.

Article Snippet: Human DPSCs were purchased from Lonza, Pharma & Biotech (Cat. # PT-5025).

Techniques: Control, Bacteria, RNA Sequencing

Journal: Frontiers in Genetics

Article Title: Gene expression profiles identify key factors in inflammatory odontoblastic dental pulp stem cell differentiation via TNFα/C5L2

doi: 10.3389/fgene.2025.1592599

Figure Lengend Snippet: Signaling pathways affected by siC5L2 in odontoblast-like differentiation of hDPSCs in dentinogenic media. hDPSCs were cultured and treated with or without siC5L2 and differentiated for 7 days. Cell lysates were collected, and RNA was prepared using the RNeasy mini kit (Qiagen), and next-generation RNA sequencing was done using the poly-A-RNA sequencing technique. Histogram showing upregulated and activated transcription factors (blue) and repressed or downregulated transcription factors (orange). Notably, TCF ( ; ; ; ; ; ), especially TCF7L2, is highly upregulated in siC5L2-stimulated odontoblast-like differentiated DPSCs.

Article Snippet: Human DPSCs were purchased from Lonza, Pharma & Biotech (Cat. # PT-5025).

Techniques: Protein-Protein interactions, Cell Culture, RNA Sequencing

Journal: Frontiers in Genetics

Article Title: Gene expression profiles identify key factors in inflammatory odontoblastic dental pulp stem cell differentiation via TNFα/C5L2

doi: 10.3389/fgene.2025.1592599

Figure Lengend Snippet: The top GO terms in the enrichment analysis are biological process, cellular component, and molecular function (MF) terms in the enrichment analysis in RNA-Seq. data. in TNFα and siC5L2 combined groups. Significantly enriched GO terms can be found in TNFα-treated C5L2-silenced hDPSCs in dentinogenic media based on their functions compared with the control. (A,B) Go-term and Reactome enrichment pathway analysis of up- and downregulated DEGs. The dot plot shows the top enriched Reactome pathways. The size of the dot is based on the gene count enriched in the pathway, and the color of the dot shows the significance of pathway enrichment. (C) Sashimi plots for quantitative visualization of RNA sequencing read alignments. The arched lines indicate splice junctions; the number of reads using that splice junction is indicated on the arched line. Data were examined on sashimi plots, which revealed the number of variants and genomic mutations on chr3, chr11, Chr10, and chr19 in TNFα-treated C5L2-silenced DPSCs in dentinogenic media against the control. Red sashimi plots show variants in the TNFα+siC5L2-treated group, and orange shows in the control. While lower black annotations are Read alignments of alternative isoforms and genomic regions of interest.

Article Snippet: Human DPSCs were purchased from Lonza, Pharma & Biotech (Cat. # PT-5025).

Techniques: RNA Sequencing, Control

Journal: Frontiers in Genetics

Article Title: Gene expression profiles identify key factors in inflammatory odontoblastic dental pulp stem cell differentiation via TNFα/C5L2

doi: 10.3389/fgene.2025.1592599

Figure Lengend Snippet: Signaling pathways affected by C5L2 silencing in odontoblast-like differentiation of hDPSCs in dentinogenic media stimulated by TNFα. hDPSCs were cultured and treated with or without TNFα (twice a week) in C5L2-silenced DPSCs and differentiated for 7 days. Cell lysates were collected, and RNA was prepared using the RNeasy mini kit (Qiagen), and next-generation RNA sequencing was done using the poly-A-RNA sequencing technique. Histogram showing upregulated and activated transcription factors (blue) and repressed or downregulated transcription factors (orange). Notably, TCF ( ; ; ; ; ), especially TCF4, is highly upregulated in C5L2-silenced TNFα-stimulated odontoblast-like differentiated DPSCs.

Article Snippet: Human DPSCs were purchased from Lonza, Pharma & Biotech (Cat. # PT-5025).

Techniques: Protein-Protein interactions, Cell Culture, RNA Sequencing

Journal: Bioactive Materials

Article Title: Dental pulp stem cells alleviate Schwann cell pyroptosis via mitochondrial transfer to enhance facial nerve regeneration

doi: 10.1016/j.bioactmat.2025.01.031

Figure Lengend Snippet: TNFα activates the NF-κB signaling pathway in DPSCs to promote mitochondrial transfer. A. Gene Ontology (GO) analysis of RNA sequencing results comparing RSC96 cells with LA-RSC96 cells (n = 3). B. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of RNA sequencing results comparing RSC96 cells with LA-RSC96 cells, with the TNFα signaling pathway highlighted in a red box. C. WB analysis of phosphorylated Iκκα/β, Iκκβ, phosphorylated p65, and total p65 expression levels in DPSCs (ATP treatment), as well as in Transwell systems involving DMSO-treated DPSCs with RSC96 cells, DMSO-treated DPSCs with LA-RSC96 cells, and TNFα-neutralizing antibody (a-TNFα)-treated DPSCs with LA-RSC96 cells. D. Quantification of protein expression levels from (C) (n = 3). E. WB analysis of NLRP3, GSDMD, pro- and cleaved-IL1β, TOM20, and β-actin as a loading control in LA-RSC96 cells and FACS-sorted LA-RSC96 cells co-cultured with DPSCs or Bay-DPSCs (Bay 11–7082, an NF-κB inhibitor-treated DPSCs). F. Quantification of protein expression levels from (E) (n = 3). G. Flow cytometry analysis of mitochondrial ROS levels in LA-RSC96 cells, and LA-RSC96 cells co-cultured with DPSCs or Bay-DPSCs. H. Quantification of relative fluorescence intensity from (G) (n = 3). I. Flow cytometry analysis of mitochondrial membrane potential in LA-RSC96 cells, and LA-RSC96 cells co-cultured with DPSCs or Bay-DPSCs. J. Quantification of relative fluorescence intensity from (I) (n = 3). K. Phalloidin-488 staining of F-actin in DPSCs treated with DMSO as control or TNFα (20 ng/mL). L. Flow cytometry analysis of mitochondrial transfer rates comparing: 1) DMSO-treated DPSCs with RSC96 cells; 2) TNFα (20 ng/mL)-treated DPSCs with RSC96 cells. M. Quantification of mitochondrial transfer rates from (L) (n = 5). Scale bars are indicated in the respective panels. Data are mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001; ns, not significant.

Article Snippet: For TNFα or Metformin preconditioning, DPSCs were treated with human TNFα (HY-P70426A, MCE) or Metformin (HY-B0627, MCE) for 24 h. Fluorescence-activated cell sorting (FACS) was performed using a FACSAria III cell sorter (BD Biosciences) to isolate RSC96 cells from the co-culture systems.

Techniques: RNA Sequencing Assay, Expressing, Control, Cell Culture, Flow Cytometry, Fluorescence, Membrane, Staining

Journal: Bioactive Materials

Article Title: Dental pulp stem cells alleviate Schwann cell pyroptosis via mitochondrial transfer to enhance facial nerve regeneration

doi: 10.1016/j.bioactmat.2025.01.031

Figure Lengend Snippet: TNFα enhances mitochondrial adaptive stress response in DPSCs, promoting mitochondrial protection against pyroptosis-induced damage in Schwann cells. A. Flow cytometry analysis of mitochondrial ROS levels in unstained DPSCs, DMSO-treated DPSCs, and TNFα (20 ng/mL)-treated DPSCs. B. Quantification of relative fluorescence intensity from (A) (n = 3). C. Flow cytometry analysis of mitochondrial membrane potential in unstained DPSCs, DMSO-treated DPSCs, and TNFα (20 ng/mL)-treated DPSCs. D. Quantification of relative fluorescence intensity from (C) (n = 3). E. ATP levels in DMSO-treated DPSCs and TNFα (20 ng/mL)-treated DPSCs (n = 3). F. WB analysis of PGC1a, NRF1, TFAM, COX4, TOM20, and β-actin as a loading control in DMSO-treated DPSCs, TNFα (20 ng/mL)-treated DPSCs, and metformin (Met, 1 nM)-treated DPSCs. G. Quantification of protein expression levels from (F) (n = 3). H. Quantitative Polymerase Chain Reaction (qPCR) analysis of Ppargc1a , Nrf1 , Tfam , mt-Co2 , Cox4i1 , and Tomm20 expression levels in DMSO-treated DPSCs and TNFα (20 ng/mL)-treated DPSCs (n = 3). I. qPCR analysis of CAT and SOD1 expression levels (n = 3). J. qPCR analysis of Pink1 and Prkn expression levels in DMSO-treated DPSCs and TNFα (20 ng/mL)-treated DPSCs (n = 3). K. WB analysis of CAT, SOD1, Parkin, and β-actin as a loading control in DMSO-treated DPSCs, TNFα (20 ng/mL)-treated DPSCs, and metformin (Met, 1 nM)-treated DPSCs. L. Quantification of protein expression levels from (K) (n = 3). M. Flow cytometry analysis of mitochondrial ROS levels in LA-RSC96 cells, and LA-RSC96 cells co-cultured with DPSCs or TNFα (20 ng/mL)-treated DPSCs. N. Quantification of relative fluorescence intensity from (M) (n = 3). O. Flow cytometry analysis of mitochondrial membrane potential in LA-RSC96 cells, and LA-RSC96 cells co-cultured with DPSCs or TNFα (20 ng/mL)-treated DPSCs. P. Quantification of relative fluorescence intensity from (O) (n = 3). Q. ATP levels in RSC96, LA-RSC96, and LA-RSC96 cells co-cultured with DPSCs or TNFα (20 ng/mL)-treated DPSCs (n = 5). R. WB analysis of NLRP3, GSDMD, pro- and cleaved-Caspase-1, pro- and cleaved-IL1β, TOM20, and β-actin as a loading control in LA-RSC96, and LA-RSC96 cells co-cultured with DPSCs or TNFα (20 ng/mL)-treated DPSCs. S. Quantification of protein expression levels from (R) (n = 3). T. qPCR analysis of IL6 , Cxcl10 , and Tnf expression levels in LA-RSC96, and LA-RSC96 cells co-cultured with DPSCs or TNFα (20 ng/mL)-treated DPSCs (n = 3). U. Caspase-1 activity assay in RSC96, LA-RSC96, and LA-RSC96 cells co-cultured with DPSCs or TNFα (20 ng/mL)-treated DPSCs (n = 5). Data are mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001; ns, not significant.

Article Snippet: For TNFα or Metformin preconditioning, DPSCs were treated with human TNFα (HY-P70426A, MCE) or Metformin (HY-B0627, MCE) for 24 h. Fluorescence-activated cell sorting (FACS) was performed using a FACSAria III cell sorter (BD Biosciences) to isolate RSC96 cells from the co-culture systems.

Techniques: Flow Cytometry, Fluorescence, Membrane, Control, Expressing, Real-time Polymerase Chain Reaction, Cell Culture, Activity Assay

Journal: Bioactive Materials

Article Title: Dental pulp stem cells alleviate Schwann cell pyroptosis via mitochondrial transfer to enhance facial nerve regeneration

doi: 10.1016/j.bioactmat.2025.01.031

Figure Lengend Snippet: TNFα preconditioning enhances the efficacy of DPSCs in repairing rat facial nerve injury. A. Facial nerve functional scores comparing the sham-operated groups, FNI, DPSC-treated groups, and TNFα-preconditioned DPSC-treated groups at different time points (n = 6). B. TEM images showing myelin sheath structures in the sham-operated groups, FNI (n = 16), DPSC-treated groups (n = 32), and TNFα-preconditioned DPSC-treated groups (n = 36). C. Quantification of G-ratio values (myelin thickness/axon diameter) based on TEM images in (B). D. WB analysis of NLRP3, GSDMD, pro- and cleaved-Caspase-1, pro- and cleaved-IL1β, and β-actin as a loading control in facial nerve tissues from the sham-operated groups, FNI, DPSC-treated groups, and TNFα-preconditioned DPSC-treated groups. E. Quantification of protein expression levels from (D) (n = 3). F. Immunofluorescence (IF) staining of GSDMD and MPZ in tissue sections from the sham-operated groups, FNI, DPSC-treated groups, and TNFα-preconditioned DPSC-treated groups. G, H. Quantification of fluorescence intensities for GSDMD and MPZ from (F) (n = 5). Scale bars are indicated in the respective panels. Data are mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001; ns, not significant.

Article Snippet: For TNFα or Metformin preconditioning, DPSCs were treated with human TNFα (HY-P70426A, MCE) or Metformin (HY-B0627, MCE) for 24 h. Fluorescence-activated cell sorting (FACS) was performed using a FACSAria III cell sorter (BD Biosciences) to isolate RSC96 cells from the co-culture systems.

Techniques: Functional Assay, Control, Expressing, Immunofluorescence, Staining, Fluorescence