Journal: Open Veterinary Journal

Article Title: Generation and characterization of mesenchymal stem cells from the affected femoral heads of dogs with Legg Calvé Perthes disease

doi: 10.5455/OVJ.2024.v14.i5.12

Figure Lengend Snippet: Trilineage differentiation potency. Canine LCPD-affected femoral head-derived adherent cells (A-C) and BMMSCs (D-F) differentiated into adipocytes (A and D), osteoblasts (B and E), and chondrocytes (C and F). (A and D): Sudan III staining, (B and E): Alizarin Red staining, and (C and F): Alcian blue staining. Scale bars = 100 μm or 1 mm.

Article Snippet: Canine osteoblast differentiation medium (Cell Applications, Inc.) was used for osteoblast differentiation, and the same static culture was performed for 2 weeks.

Techniques: Derivative Assay, Staining

Journal: Communications Biology

Article Title: mTORC1 impedes osteoclast differentiation via calcineurin and NFATc1

doi: 10.1038/s42003-018-0028-4

Figure Lengend Snippet: Late mTORC1 deletion in myeloid lineage enhances osteoclastogenesis. Analyses of Raptor fl/fl -Lyz-Cre cKO mice and littermate controls. a , b Expression of osteoclast markers ( a ) and osteoclastogenic transcription factors ( b ) in bone marrow osteoclast differentiation cultures ( n = 6). c Representative images of TRAP-stained osteoclast differentiation cultures. d Osteoclast resorptive activity measured by calcium release from bone plates ( n = 16). e Osteoclast precursor proliferation by BrdU incorporation ( n = 10). f , g mTORC1 signaling in bone marrow osteoclast differentiation cultures from Raptor fl/fl -Lyz mice or littermate controls 50 h after RANKL treatment. f Phosphorylation of mTOR; g Phosphorylation of S6K1. p/T, ratio of phosphorylated/total protein. h – t Bone phenotype in 3-month-old male mice. h Serum CTX-1 bone resorption marker ( n = 8). i Serum P1NP bone formation marker ( n = 8). j Representative μCT images of the trabecular bone of the tibial metaphysis (top) and the entire proximal tibia (bottom). k – p Quantification of trabecular bone volume and architecture in proximal tibiae by μCT ( n = 8–10). k BV/TV bone volume/tissue volume ratio; l BS bone surface; m Tb.N trabecular number; n Conn. D. connectivity density; o Tb.Sp trabecular separation; p SMI structure model index. q – t Bone histomorphometry of distal femurs ( n = 6). Oc.S osteoclast surface, B.S bone surface, Oc.N osteoclast number, Ob.S osteoblast surface, Ob.N osteoblast number. Error bars, SEM; *, p < 0.05; **, p < 0.01; ***, p < 0.005; ****, p < 0.001; n.s. non-significant. R raptor. Full-size scans of immunoblots are shown in Supplementary Fig.

Article Snippet: Osteoblast precursors were expanded for 4 days in Mesenchymal Stem Cell (MSC) media using a Mouse MesenCult Proliferation Kit (StemCell Technologies) before the addition of osteoblast differentiation cocktail (StemCell Technologies).

Techniques: Expressing, Staining, Activity Assay, BrdU Incorporation Assay, Phospho-proteomics, Marker, Western Blot

Journal: Communications Biology

Article Title: mTORC1 impedes osteoclast differentiation via calcineurin and NFATc1

doi: 10.1038/s42003-018-0028-4

Figure Lengend Snippet: Constitutive mTORC1 activation in myeloid lineage impedes osteoclast differentiation. Analyses of Tsc1 fl/fl -Lyz-Cre cKO mice and littermate controls. a , b Expression of osteoclast markers ( a ) and osteoclastogenic transcription factors ( b ) in bone marrow osteoclast differentiation cultures ( n = 6). c Representative images of TRAP-stained osteoclast differentiation cultures. d Osteoclast resorptive activity measured by calcium release from bone plates ( n = 16). e Osteoclast precursor proliferation by BrdU incorporation ( n = 10). f , g mTORC1 signaling in bone marrow osteoclast differentiation cultures 60 h after RANKL treatment. f Phosphorylation of mTOR; g Phosphorylation of S6K1. p/T ratio of phosphorylated/total protein. h – r Bone phenotype in 2-month-old male mice. h Serum CTX-1 bone resorption marker ( n = 8). i Serum P1NP bone formation marker ( n = 8). j Bone histomorphometry of distal femurs ( n = 6). Oc.S osteoclast surface, B.S bone surface, Oc.N osteoclast number. k Bone histomorphometry of distal femurs ( n = 6). Ob.S osteoblast surface, B.S bone surface, Ob.N osteoblast number. l Representative μCT images of the trabecular bone of the tibial metaphysis (top) and the entire proximal tibia (bottom). m – r Quantification of trabecular bone volume and architecture in proximal tibiae by μCT ( n = 8). m BV/TV bone volume/tissue volume ratio, n BS bone surface, o Tb.N trabecular number, p Tb.Sp trabecular separation, q Conn. D. connectivity density, r SMI structure model index. Error bars, SEM; * p < 0.05; ** p < 0.01; *** p < 0.005; **** p < 0.001; n.s. non-significant. T Tsc1. Full-size scans of immunoblots are shown in Supplementary Fig.

Article Snippet: Osteoblast precursors were expanded for 4 days in Mesenchymal Stem Cell (MSC) media using a Mouse MesenCult Proliferation Kit (StemCell Technologies) before the addition of osteoblast differentiation cocktail (StemCell Technologies).

Techniques: Activation Assay, Expressing, Staining, Activity Assay, BrdU Incorporation Assay, Phospho-proteomics, Marker, Western Blot