Journal: Stem Cells International

Article Title: Proteomic Analysis Reveals Commonly Secreted Proteins of Mesenchymal Stem Cells Derived from Bone Marrow, Adipose Tissue, and Synovial Membrane to Show Potential for Cartilage Regeneration in Knee Osteoarthritis

doi: 10.1155/2021/6694299

Figure Lengend Snippet: Representative characterization of bone marrow (BM), adipose tissue (AT), and synovial membrane (SM) mesenchymal stem cells (MSCs). (a) Phase-contrast microscopy images of MSCs (scale bar = 100 μ m). (b) Differentiation potential of MSCs. Adipogenic differentiation (Oil Red O, top), osteogenic differentiation (Alizarin Red S, middle), and chondrogenic differentiation (Alcian Blue, bottom). (c) The immunophenotyping of MSCs using flow cytometry. Positive (CD73, CD90, and CD105) and negative (CD14, CD34, CD45, CD79a, and human leukocyte antigen-DR (HLA-DR)) markers. The histogram is shown with an overlay isotype control.

Article Snippet: After 2 days, differentiation was induced using chondrogenic differentiation medium (containing Dulbecco's minimal EM-high glucose (HyClone), 10% FBS, 1% P/S, 0.1 μ M of dexamethasone, 1X insulin-transferrin-selenium (Gibco), 50 μ M of L-ascorbic acid 2-phosphate sesquimagnesium salt hydrate, and 5 ng/mL of transforming growth factor β 1 (TGF- β 1; PeproTech, Rocky Hill, NJ, USA)) [ ].

Techniques: Microscopy, Flow Cytometry

Journal: bioRxiv

Article Title: Cellular and molecular changes in the human osteoarthritic and aging hip pulvinar

doi: 10.1101/2024.04.21.590119

Figure Lengend Snippet: Freshly obtained pulvinar tissues were enzymatically dissociated and viable cells were seeded in 24-well plates at clonal density in human Mesencult medium. A) Colony forming unit-fibroblast (CFU-f) assays were performed at various initial seeding densities, but CFU-f’s were only observed starting at 2 and 5×10 4 cells/cm 2 . An initial seeding density of 5×10 5 cells/cm 2 was used in subsequent assays. CFU-f’s were never observed when samples from non-osteoarthritic patients were analyzed (G1 and G2, n=6 and 13 respectively, for all seeding densities tested). B) CFU-f’s from G3 and G4 samples were expanded by passaging at low density until they reached senescence. The primary MSPC cell lines could be expanded for an average of nine passages. C) Multilineage differentiation assays of passage 3 MSPCs obtained from G3 and G4 pulvinars show that each primary cell line tested possessed adipogenic, osteogenic and chondrogenic cells potential (n= 5 and n= 6 respectively for G3 and G4).

Article Snippet: For the chondrogenic differentiation, cells were pelleted in Chondrogenic differentiation media (StemCell Technologies) and media was replaced as recommended for 21 days followed by fixation and staining with Sox9, DAPI and Collagen I.

Techniques: Passaging

Journal: The Scientific World Journal

Article Title: In Vitro Chondrogenesis Transformation Study of Mouse Dental Pulp Stem Cells

doi: 10.1100/2012/827149

Figure Lengend Snippet: Characteristics of chondrocyte derived from mouse DPSC. After chondrogenic induction, the cytoplasm contracted toward the nucleus and formed spherical cells without branch (a and b). Glycosaminoglycans in chondrocyte tissue were stained by toluidine blue and showed blue in colors (c). Control groups without chondrogenic induction (d).

Article Snippet: After washing the cells with 1 X PBS, the cells were placed in chondrogenic medium (Zen-Bio, Inc.) for 21 days to observe for chondrocyte differentiation.

Techniques: Derivative Assay, Staining, Control

Journal: International Journal of Molecular Medicine

Article Title: SETD7 regulates chondrocyte differentiation and glycolysis via the Hippo signaling pathway and HIF-1α

doi: 10.3892/ijmm.2021.5043

Figure Lengend Snippet: mRNA expression of SETD7 decreases during chondrogenic differentiation. (A) Alcian blue staining of differentiated ATDC5 cells cultured in micromass at 1 and 20% O 2 . Scale bar, 100 µ m. mRNA expression of (B) aggrecan, (C) SOX9, (D) COL2A1 and (E) SETD7 during chondrogenic differentiation at 1% O 2 . Results are representative of ≥3 independent experiments. * P<0.05, *** P<0.001 vs. 0 days. SETD7, SET domain containing 7; HIF-1α, hypoxia inducible factor-1α; COL2A1, collagen II, α1; SOX9, SRY-related box gene 9.

Article Snippet: At 70-90% confluence, growth medium was replaced with chondrogenic differentiation medium supplemented with 1x insulin-transferrin-selenium (Corning, Inc.).

Techniques: Expressing, Staining, Cell Culture

Journal: International Journal of Molecular Medicine

Article Title: SETD7 regulates chondrocyte differentiation and glycolysis via the Hippo signaling pathway and HIF-1α

doi: 10.3892/ijmm.2021.5043

Figure Lengend Snippet: Knocking down SETD7 activates chondrogenic differentiation. (A) Immunofluorescence staining of ATDC5 cells at 1% O 2 incubated with secondary antibodies and DAPI. (B) Immunofluorescence staining with SETD7 primary antibody, secondary antibodies and DAPI in ATDC5 cells at 1% O 2 . (C) mRNA expression and (D) protein levels of SETD7 after knocking down SETD7 at 1% O 2 . (E) Alcian blue staining after knocking down SETD7 in cells cultured in micromass at 1% O 2 . mRNA expression of (F) aggrecan, (G) SOX9 and (H) COL2A1 after knocking down SETD7 at 1% O 2 . * P<0.05, ** P<0.01, *** P<0.001 vs. con. SETD7, SET domain containing 7; HIF-1α, hypoxia inducible factor-1α; COL2A1, collagen II, α1; SOX9, SRY-related box gene 9; con, control; sh, short hairpin.

Article Snippet: At 70-90% confluence, growth medium was replaced with chondrogenic differentiation medium supplemented with 1x insulin-transferrin-selenium (Corning, Inc.).

Techniques: Immunofluorescence, Staining, Incubation, Expressing, Cell Culture, Control

Journal: International Journal of Molecular Medicine

Article Title: SETD7 regulates chondrocyte differentiation and glycolysis via the Hippo signaling pathway and HIF-1α

doi: 10.3892/ijmm.2021.5043

Figure Lengend Snippet: SETD7 regulates chondrocyte differentiation and glycolysis via the Hippo and HIF-1α. In normal conditions, SETD7 activates the Hippo signaling pathway, which phosphorylates YAP and retains it in the cytoplasm. In hypoxic conditions, expression of SETD7 is inhibited, resulting in increased YAP and HIF-1α in the cytoplasm. The accumulated YAP and HIF-1α translocate into the nucleus and combine to form a complex, which further promotes expression of glycolysis-associated genes and chondrogenic differentiation. SETD7, SET domain containing 7; YAP, Yes-associated protein; HIF-1α, hypoxia inducible factor-1α; GLUT1, glucose transporter 1; LDHA, lactate dehydrogenase A; PGK1, phosphoglycerate kinase 1; PKM2, pyruvate kinase isoenzyme 2; LATS1, large tumor suppressor 1; SOX9, SRY-related box gene 9; Me, methylation; P, phosphorylation.

Article Snippet: At 70-90% confluence, growth medium was replaced with chondrogenic differentiation medium supplemented with 1x insulin-transferrin-selenium (Corning, Inc.).

Techniques: Expressing, Methylation, Phospho-proteomics