type ii collagen immunohistochemistry  (Developmental Studies Hybridoma Bank)

Journal: eLife

Article Title: Identification of a sub-population of synovial mesenchymal stem cells with enhanced treatment efficacy in a rat model of osteoarthritis

doi: 10.7554/eLife.103332

Figure Lengend Snippet: Representative images of a rat with DMM surgery ( A ) with MSC injection (TdTomato – B) and collagen 2 staining ( C ). Scale bars equal 20 µm.

Article Snippet: Antigen retrieval was achieved using 10 mM sodium citrate (pH 6.0), and non-specific blocking was prevented using goat serum (1:500 dilution in TBST). tdTomato (AB8181, Origene), Collagen 2 (Col2; Clone # II-II6B3, DSHB), PRG4 (Clone # 9G3, Millipore), Ccl2 (Clone # 2D8, Thermo Fisher), Ki67 (Clone # SolA15, ThermoFisher), Sox9 (Clone # 7H13L8, Thermo Fisher) or CD47 (Clone # B6H12, Thermo Fisher) were applied to the sections and incubated overnight.

Techniques: Injection, Staining

Journal: eLife

Article Title: Identification of a sub-population of synovial mesenchymal stem cells with enhanced treatment efficacy in a rat model of osteoarthritis

doi: 10.7554/eLife.103332

Figure Lengend Snippet: The gating strategy to identify and sort CD47 Hi vs. CD47 Lo cell populations ( A–C ). CD47 Hi vs. CD47 Lo cells were identified and isolated from normal and osteoarthritis (OA) synovial tissue ( B ). Chondrogenic differentiation of CD47 Hi vs. CD47 Lo cells using pellet culture and stained with Alcian Blue ( D ). GAG quantification of the pellets ( E ). The pellets were also stained with Collagen 2 (Col2) as a marker of mature cartilage ECM ( E, G ). Signifiance determined by1 way ANOVA. p <0.05. Scale bars equal 100 µm.

Article Snippet: Antigen retrieval was achieved using 10 mM sodium citrate (pH 6.0), and non-specific blocking was prevented using goat serum (1:500 dilution in TBST). tdTomato (AB8181, Origene), Collagen 2 (Col2; Clone # II-II6B3, DSHB), PRG4 (Clone # 9G3, Millipore), Ccl2 (Clone # 2D8, Thermo Fisher), Ki67 (Clone # SolA15, ThermoFisher), Sox9 (Clone # 7H13L8, Thermo Fisher) or CD47 (Clone # B6H12, Thermo Fisher) were applied to the sections and incubated overnight.

Techniques: Isolation, Staining, Marker

Journal: bioRxiv

Article Title: Nascent extracellular matrix converts biomaterial cues into cell fate decisions

doi: 10.64898/2026.01.10.698826

Figure Lengend Snippet: a. Representative fluorescent images of nuclei and cell membrane of chondrocytes cultured in low, mid, and high modification hydrogels at day 7 (scale bars: 10 μm.) b. Quantification of divided nuclei of chondrocytes culture in low, mid and high modification hydrogels at day1, 4 and 7 (low: day 1 n = 12 regions of interest (ROIs), N = 2; day 4 n = 9 ROIs, N = 2; day 7 n = 12 ROIs, N = 3; mid: day 1 n = 8 ROIs, N = 2; day 4 n = 8 ROIs, N = 2; day 7 n = 12 ROIs, N = 3; high: day 1 n = 10 ROIs, N = 2; day 4 n = 9 ROIs, N = 2; day 7 n = 12 ROIs, N = 3) c. Quantification of divided nuclei of chondrocytes culture in low, mid and high modification hydrogels at day 7 ( low: n = 12 ROIs, N = 3; mid: n = 12 ROIs, N = 3; and high: n = 12 ROIs, N = 3). d. Representative fluorescent images of incorporation of 5-ethynyl-2’-deoxyuridine (EdU) into chondrocytes cultured in low, mid and high modification hydrogels at day 7 (scale bar: 100 μm). e. Quantification of EdU incorporation into chondrocytes cultured in low, mid and high modification hydrogels at day 7 (scale bar: 100 μm, low: n = 23 ROIs, N = 4; mid: n = 24 ROIs, N = 4; and high: n = 23 ROIs, N = 4 ). f. Quantification of chondrocyte cell aspect ratio at day 7 (low: n = 108 cells, N = 3; mid: n = 112 cells, N = 3; and high: n = 111 cells, N = 3). g. Schematic showing the downstream effects of Sox9 translocation into the nucleus, including upregulation of collagen type IIA1, aggrecan (ACAN) and Sox9, and the downregulation of collagen type IA1, versican (VCAN) and THY-1 h. Representative immunofluorescent images and quantification of Sox9 nuclear translocation calculated by the nucleus-to-cytoplasm (NC) ratio at day 7 (scale bar: 10 μm, low: n = 139 cells, N = 3; and high: n = 124 cells, N = 3). i. Ratio of COL2A1/COL1A1 and ACAN/VCAN gene expression at day 7, measured by qPCR and normalized to S18 housekeeping gene. Gene expression ratios were calculated as 2 (-(ΔCt₁ - ΔCt₂)) , where ΔCt₁ and ΔCt₂ represent Ct values of each gene normalized to S18. ( N = 4) a-i. N = number of independent experiments, error bar = standard deviation, ****p < 0.0001, **p < 0.01, *p < 0.05, ns: not significant by one-way ANOVA with Tukey’s multiple comparisons test.

Article Snippet: Primary antibodies included SOX9 (NovusBio, NBP2-24659, 1:100), Collagen II (DSHB, II-II6B3, 1:100), Collagen VI (Biosynth, 70R-CR009X, 1:100), Decorin (Kerafast, ENH077-FP, 1:100), and Aggrecan (Abcam, ab3778, 1:50).

Techniques: Membrane, Cell Culture, Modification, Translocation Assay, Gene Expression, Standard Deviation

Journal: bioRxiv

Article Title: Nascent extracellular matrix converts biomaterial cues into cell fate decisions

doi: 10.64898/2026.01.10.698826

Figure Lengend Snippet: a. Timeline and schematic showing the blocking of cell adhesion to hyaluronic acid with CD44 function-perturbing antibody for 1 hour pre-embedding and for 3 consecutive days while maintaining adhesion to nECM. b. Representative fluorescent images and quantification of the incorporation of EdU in chondrocytes cultured without (Ctrl) and with CD44 inhibition (CD44i) in ‘low’ modification hydrogels at day 7. (scale bar = 100μm, low-Ctrl: n = 23 ROIs, N = 3; low-CD44: n = 27 ROIs, N = 3; high-Ctrl: n = 23 ROIs, N = 3) c. Representative fluorescent images (dashed line outlines nuclei) and quantification of Sox9 nucleus-to-cytoplasm (NC) ratio of chondrocytes cultured without (Ctrl) and with CD44 inhibition (CD44i) in low modification hydrogels at day 7. (scale bar = 10μm, low Ctrl: n = 138 cells, N = 4; low CD44i: n = 110 cells; N = 4, high-Ctrl: n =124 cells; N = 4) a-c. N = number of independent experiments, error bar = standard deviation, ****p < 0.0001, ns: not significant by one-way ANOVA with Tukey’s multiple comparisons test.

Article Snippet: Primary antibodies included SOX9 (NovusBio, NBP2-24659, 1:100), Collagen II (DSHB, II-II6B3, 1:100), Collagen VI (Biosynth, 70R-CR009X, 1:100), Decorin (Kerafast, ENH077-FP, 1:100), and Aggrecan (Abcam, ab3778, 1:50).

Techniques: Blocking Assay, Cell Culture, Inhibition, Modification, Standard Deviation

Journal: bioRxiv

Article Title: Nascent extracellular matrix converts biomaterial cues into cell fate decisions

doi: 10.64898/2026.01.10.698826

Figure Lengend Snippet: a. Timeline and schematic showing the blocking of cell adhesion to nECM using an integrin β1 (ITGB1) function perturbing antibody for 1 hour pre-embedding and for 7 consecutive days . b. Representative fluorescent images of the incorporation of EdU in chondrocytes cultured without (Ctrl) and with ITGB1 inhibition (ITGB1i) in high and low modification hydrogels at day 7 (scale bar = 100 μm). c. Quantification of the incorporation of EdU in chondrocytes cultured without (Ctrl) and with ITGB1 inhibition (ITGB1i) in high and low modification hydrogels at day 7 (low-Ctrl: n = 23 ROIs, N = 3; low-ITGB1i: n = 30 ROIs, N = 3; high-Ctrl: n = 23 ROIs, N = 3; low-ITGB1i: n = 27 ROIs, N = 3). d. Representative fluorescent images of Sox9 immunofluorescence (dashed line outlines nuclei) of chondrocytes cultured without (Ctrl) and with ITGB1 inhibition (ITGB1i) in low and high modification hydrogels at day 7 (scale bar = 10 μm). e. Quantification of Sox9 nucleus-to-cytoplasm (NC) ratio of chondrocytes cultured without (Ctrl) and with ITGB1 inhibition (ITGB1i) in low and high modification hydrogels at day 7 (low-Ctrl: n = 138 cells, N = 4; low-ITGB1i: n = 133 cells, N = 4; high-Ctrl: n = 124 cells, N = 4; high-ITGBi: n = 138 cells, N = 4). a-e. N = number of independent experiments ****p < 0.0001, *p < 0.05, error bar = standard deviation, ns: no significant difference by two-way ANOVA with Tukey’s multiple comparisons test.

Article Snippet: Primary antibodies included SOX9 (NovusBio, NBP2-24659, 1:100), Collagen II (DSHB, II-II6B3, 1:100), Collagen VI (Biosynth, 70R-CR009X, 1:100), Decorin (Kerafast, ENH077-FP, 1:100), and Aggrecan (Abcam, ab3778, 1:50).

Techniques: Blocking Assay, Cell Culture, Inhibition, Modification, Immunofluorescence, Standard Deviation