Journal: Journal of Tissue Engineering

Article Title: Synergistic Wnt/BMP Co-activation accelerates osteogenic differentiation of human pluripotent stem cells via paraxial mesoderm induction

doi: 10.1177/20417314261468879

Figure Lengend Snippet: A schematic showing BMP4-mediated osteogenesis maturation through CHIR-induced mesodermal differentiation through Wnt activation and BMP signaling, efficient osteoblast differentiation with hiPSCs is synergistically promoted through Wnt/BMP signaling. hiPSC, Human induced pluripotent stem cells; CHIR, CHIR99021; BMP4, Bone morphogenetic protein 4.

Article Snippet: The medium consisted of STEMdiffTM APELTM2 Medium (APEL; 05270, STEMCELL, Canada) supplemented with 6 ng/mL StemMACS CHIR99021 (CHIR; 130-103-926, Miltenyi Biotec, Germany) and 50 ng/mL BMP4 (CYT-1093, ProSpec, Korea) for two days.

Techniques: Activation Assay

Journal: Journal of Tissue Engineering

Article Title: Synergistic Wnt/BMP Co-activation accelerates osteogenic differentiation of human pluripotent stem cells via paraxial mesoderm induction

doi: 10.1177/20417314261468879

Figure Lengend Snippet: Coactivation of Wnt/BMP signaling specifies the differentiation of hiPSCs to a paraxial mesoderm. (a) A schematic diagram of the protocol of mesoderm according to the differentiation period. (b-e) Graphs of mRNA expression of MSGN1 , TBX6 , ISL1 , and NKX2.5 in mesenchymal-induced cultures (n=3). (f-g) Graphs of MSGN1 and RUNX2 mRNA expression according to the concentrations of BMP2 and BMP4 (n=3 and 5). (h-i) Graphs of TBX6 and RUNX2 mRNA expression for BMP4 (n=5). Groups: PM, Pluripotent stem cell Medium; MM, Mesoderm differentiation Medium; AC, CHIR; ACB, CHIR+BMP4. hiPSC, Human induced pluripotent stem cells; CHIR, CHIR99021; BMP2, Bone morphogenetic protein 2; BMP4, Bone morphogenetic protein 4. All data represent mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.

Article Snippet: The medium consisted of STEMdiffTM APELTM2 Medium (APEL; 05270, STEMCELL, Canada) supplemented with 6 ng/mL StemMACS CHIR99021 (CHIR; 130-103-926, Miltenyi Biotec, Germany) and 50 ng/mL BMP4 (CYT-1093, ProSpec, Korea) for two days.

Techniques: Expressing

Journal: Journal of Tissue Engineering

Article Title: Synergistic Wnt/BMP Co-activation accelerates osteogenic differentiation of human pluripotent stem cells via paraxial mesoderm induction

doi: 10.1177/20417314261468879

Figure Lengend Snippet: Total RNA Sequencing. (a) Transcription profile of 75 genes relevant to osteogenesis for hiPSC cultured for 2 days in mesoderm differentiation medium. Relative expression: red (high) and blue (low) (b) Heatmaps for markers related to osteogenesis. Relative expression: red (high) and green (low) (c) Gene ontology enrichment dot plot. Relative expression: red (high) and big dot (high). Groups: AC, CHIR; ACB, CHIR+BMP4. hiPSC, Human induced pluripotent stem cells.

Article Snippet: The medium consisted of STEMdiffTM APELTM2 Medium (APEL; 05270, STEMCELL, Canada) supplemented with 6 ng/mL StemMACS CHIR99021 (CHIR; 130-103-926, Miltenyi Biotec, Germany) and 50 ng/mL BMP4 (CYT-1093, ProSpec, Korea) for two days.

Techniques: RNA Sequencing, Cell Culture, Expressing

Journal: Journal of Tissue Engineering

Article Title: Synergistic Wnt/BMP Co-activation accelerates osteogenic differentiation of human pluripotent stem cells via paraxial mesoderm induction

doi: 10.1177/20417314261468879

Figure Lengend Snippet: Limited activation of the BMP signaling in the mesoderm period promotes the maturation of osteoblasts. (a) Schematic protocol for osteoblast differentiation via mesoderm. (b) Optical images of hiPSC according to differentiation (X40). (c-d) Graphs of mRNA gene expression of RUNX2 and COL1 according to BMP4 concentration. (e-g) Graphs of mRNA expression of COL1 , ON , and OCN . ON: osteonectin, OCN: osteocalcin. Scale bar: 500 μm. Groups: DAG, Dorsal axial mesoderm; APEL, Albumin, polyvinyl alcohol, essential lipids; AC, CHIR; ACB, CHIR+BMP4. hiPSC, Human induced pluripotent stem cells; CHIR, CHIR99021; BMP4, Bone morphogenetic protein 4. All data represent mean ± SD (n=5). *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.

Article Snippet: The medium consisted of STEMdiffTM APELTM2 Medium (APEL; 05270, STEMCELL, Canada) supplemented with 6 ng/mL StemMACS CHIR99021 (CHIR; 130-103-926, Miltenyi Biotec, Germany) and 50 ng/mL BMP4 (CYT-1093, ProSpec, Korea) for two days.

Techniques: Activation Assay, Gene Expression, Concentration Assay, Expressing

Journal: Journal of Tissue Engineering

Article Title: Synergistic Wnt/BMP Co-activation accelerates osteogenic differentiation of human pluripotent stem cells via paraxial mesoderm induction

doi: 10.1177/20417314261468879

Figure Lengend Snippet: Coactivation of Wnt/BMP signaling facilitates the maturation of osteoblasts. (a) Optical images of hiPSC for 9 days with or without BMP4 (X40). (b) ARS images of hiPSC on day 9 (X40). (c) Quantification graph of ARS absorbance at 540 nm wavelength (n=4). (d) Von Kossa images of hiPSC on day 9 (X100). (e) Quantification graph of Von Kossa + area (n=6). (f) Immunofluorescence staining images for hiPSC after osteogenic differentiation (X200). (g) Quantification graph of RUNX2 + and COL1 + area (n=3). Scale bars: (a-b) 500 μm, (d) 200 μm, (f) 100 μm. Groups: AC, CHIR; ACB, CHIR+BMP4. All data represent mean ± SD. *P < 0.05, **P < 0.01, and ****P < 0.0001. The symbol * indicates comparisons with the AC group.

Article Snippet: The medium consisted of STEMdiffTM APELTM2 Medium (APEL; 05270, STEMCELL, Canada) supplemented with 6 ng/mL StemMACS CHIR99021 (CHIR; 130-103-926, Miltenyi Biotec, Germany) and 50 ng/mL BMP4 (CYT-1093, ProSpec, Korea) for two days.

Techniques: Immunofluorescence, Staining

Journal: Journal of Tissue Engineering

Article Title: Synergistic Wnt/BMP Co-activation accelerates osteogenic differentiation of human pluripotent stem cells via paraxial mesoderm induction

doi: 10.1177/20417314261468879

Figure Lengend Snippet: Differentiation of bone formation in vivo . (a) A schematic diagram of in vivo experiments. (b) Graph of mouse weight change. (c) Images of scaffold incubated for 6 weeks in vivo . (d) 3D rendering images for micro-CT image of scaffold. (e) Images of H&E on scaffold (X200). (f) Images of von Kossa staining on the scaffold (X200). Scale bars: (c) 50 mm, (d) 5 mm, (e-f) 100 μm. Groups: Control, no treatment; Scaffold, scaffold only; BMSC, Scaffold+BMSC; AC, Scaffold+BMSC+CHIR; ACB, Scaffold+BMSC+CHIR+BMP4. All data represent mean ± SD (n=3). *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. The symbol * indicates comparisons with the Scaffold group.

Article Snippet: The medium consisted of STEMdiffTM APELTM2 Medium (APEL; 05270, STEMCELL, Canada) supplemented with 6 ng/mL StemMACS CHIR99021 (CHIR; 130-103-926, Miltenyi Biotec, Germany) and 50 ng/mL BMP4 (CYT-1093, ProSpec, Korea) for two days.

Techniques: In Vivo, Incubation, Micro-CT, Staining, Control