chondrogenic differentiation induction medium  (Novus Biologicals)

Journal: Frontiers in Cell and Developmental Biology

Article Title: Comparative cardiomyocyte differentiation potential of rat adipose-derived mesenchymal stem cells from two anatomical sites: metabolomic profiling and pathway analysis

doi: 10.3389/fcell.2025.1604605

Figure Lengend Snippet: Multilineage differentiation potential of AD-MSCs from peri-ovarian and peri-renal adipose tissue. Light microscopy images demonstrate the differentiation of third-passage AD-MSCs into osteogenic, chondrogenic, and adipogenic lineages under specific induction conditions. Scale bar: 100 μm.

Article Snippet: Third-passage AD-MSCs were plated at a density of 1 × 10 5 cells/well in 6-well plates and maintained in serum-free chondrogenic induction medium (PromoCell C-28012, Heidelberg, Germany).

Techniques: Light Microscopy

Journal: Science Advances

Article Title: Identification of human cranio-maxillofacial skeletal stem cells for mandibular development

doi: 10.1126/sciadv.ado7852

Figure Lengend Snippet: UMAP atlas of integrated analysis from cellular lineages (common progenitors, osteogenic cells, and chondrogenic cells) of human embryonic mandibles and mesenchymal cells of ( A ) human 8-WPC calvaria, ( B ) human 8-WPC long bone, ( C ) human GW9 vertebra, and ( D ) mouse E10.5 to E14.5 mandibles. CMSSC-like cells were circled with a dashed line and feature plots to the right visualize expression of DLX5 , RUNX2 , and IFITM5 . H&E staining of ( E ) human GW9 calvaria, ( F ) human GW9 long bone, ( G ) human GW9 vertebra, and ( H ) mouse E14.5 and E16.5 mandibles. Immunofluorescence staining of RUNX2 or IFITM5 of the area in the dotted box. Scale bars, 100 μm.

Article Snippet: For chondrogenic differentiation, 3 × 10 5 cells were pelleted and fed with a chondrogenic differentiation induction medium (Cyagen Biosciences, HUXMX-90041) for 27 days, with medium changes every 3 days.

Techniques: Expressing, Staining, Immunofluorescence

Journal: Science Advances

Article Title: Identification of human cranio-maxillofacial skeletal stem cells for mandibular development

doi: 10.1126/sciadv.ado7852

Figure Lengend Snippet: ( A ) Experimental flowchart of IFITM5 + cells isolation and characterization in vitro. ( B ) Flow cytometry gating strategies for sorting IFITM5 + cells ( n = 3 embryos). ( C ) Flow cytometry plots showing serial colony formation from a single IFITM5 + cell ( n = 3 clones). ( D ) Representative crystal violet staining of fibroblast colony-forming unit (CFU-F) colonies from IFITM5 − and IFITM5 + cells. ( E ) Numbers and mean diameters of CFU-F colonies ( n = 3 embryos). ( F , H , and J ) Representative alizarin red, alcian blue, and oil red O stainings after in vitro differentiation of clonally expanded IFITM5 − and IFITM5 + cells. ( G , I , and K ) qPCR analyses of osteogenic, chondrogenic, and adipogenic marker genes and quantification of alizarin red in clonally expanded IFITM5 − and IFITM5 + cells after in vitro differentiation ( n = 3 embryos). ( L ) Workflow of IFITM5 + cell characterization in vivo. ( M ) Bright-field images and H&E staining of subcapsular xenografts of IFITM5 − and IFITM5 + cells ( n = 5). ( N , P , and Q ) Immunofluorescence staining of COLII, COLI, OPN, and Stem101. ( O ) Micro–computed tomography (CT) three-dimensional (3D) reconstruction images (top) and coronal images (bottom) of mandibular defect repair after GelMA, IFITM5 − , and IFITM5 + cell transplantation. ( R ) Quantification of bone formation parameters at defect region ( n = 5). ( S ) H&E staining (bone defect edge demarcated by black dashed lines) and Masson staining in coronal sections of GelMA, IFITM5 − , and IFITM5 + cell groups. ( T ) Immunohistochemical staining for human nucleoli, RUNX2, and OPN in areas marked by white dashed lines in Masson staining images. Scale bars, 100 μm in all figures. [(E), (G), (I), and (K)] ** P < 0.01 and *** P < 0.001 determined by an unpaired two-tailed Student’s t test. (R) * P < 0.05, ** P < 0.01, and *** P < 0.001 versus GelMA; ### P < 0.001 versus IFITM5 − cells determined by one-way analysis of variance (ANOVA) with Tukey’s post hoc test. SSC-A, side scatter-area; OD, optical density.

Article Snippet: For chondrogenic differentiation, 3 × 10 5 cells were pelleted and fed with a chondrogenic differentiation induction medium (Cyagen Biosciences, HUXMX-90041) for 27 days, with medium changes every 3 days.

Techniques: Isolation, In Vitro, Flow Cytometry, Clone Assay, Staining, Marker, In Vivo, Immunofluorescence, Micro-CT, Transplantation Assay, Immunohistochemical staining, Two Tailed Test

Journal: Science Advances

Article Title: Identification of human cranio-maxillofacial skeletal stem cells for mandibular development

doi: 10.1126/sciadv.ado7852

Figure Lengend Snippet: ( A ) Schematic diagram of characterization IFITM5 + cells in vivo and in vitro. ( B ) H&E and immunofluorescence staining of IFITM5 and RUNX2 in mandible fragments aged 19, 32, and 49 (white boxes indicating magnified regions). y, years. ( C ) Flow analysis with MSC-specific surface markers and morphology of BMSCs ( n = 3). ( D ) Gating scheme for sorting Zombie − IFITM5 + BMSCs ( n = 3 patients). ( E ) Crystal violet staining of CFU-F colonies from IFITM5 − and IFITM5 + BMSCs. ( F ) Numbers and mean diameters of CFU-F colonies ( n = 3 patients). ( G , J , and L ) Alizarin red, alcian blue, and oil red O stainings after in vitro differentiation. ( H and M ) Quantification of alizarin red and oil red O stainings. ( I , K , and N ) qPCR analyses of osteogenic, chondrogenic, and adipogenic marker genes ( n = 3 patients). ( O ) Bright-field images and H&E staining of subcapsular xenografts of IFITM5 − and IFITM5 + BMSCs ( n = 5). ( P , Q , and R ) Immunofluorescence staining images of COLII, COLI, OPN, and Stem101. ( S ) Micro-CT 3D reconstruction images and coronal images of mandibular defect after GelMA, IFITM5 − , and IFITM5 + BMSC transplantation. ( T ) Quantification of bone formation parameters at defect region ( n = 5). ( U ) H&E staining (bone defect edge demarcated by black dashed lines) and Masson staining in coronal sections of GelMA, IFITM5 − , and IFITM5 + BMSC groups. ( V ) Immunohistochemical staining for Human nucleoli, RUNX2, and OPN in areas marked by white dashed lines in Masson staining images. Scale bars, 100 μm in all figures. [(F), (H), (I), (K), (M), and (N)] ** P < 0.01 and *** P < 0.001 determined by an unpaired two-tailed Student’s t test. (T) ** P < 0.01 and *** P < 0.001 versus GelMA; ## P < 0.01 and ### P < 0.001 versus IFITM5 − cells determined by one-way ANOVA with Tukey’s post hoc test.

Article Snippet: For chondrogenic differentiation, 3 × 10 5 cells were pelleted and fed with a chondrogenic differentiation induction medium (Cyagen Biosciences, HUXMX-90041) for 27 days, with medium changes every 3 days.

Techniques: In Vivo, In Vitro, Immunofluorescence, Staining, Marker, Micro-CT, Transplantation Assay, Immunohistochemical staining, Two Tailed Test

Journal: International Journal of Molecular Sciences

Article Title: In Vitro Modelling of Osteogenesis Imperfecta with Patient-Derived Induced Mesenchymal Stem Cells

doi: 10.3390/ijms25063417

Figure Lengend Snippet: Validation of induced chondrocyte phenotype. ( a ) Chondrogenic differentiation of induced mesenchymal stem cells (iMSC) lines was analyzed with Alcian blue; blue staining indicates sulfated proteoglycans deposits (scale bar size 100 µm). Representative images for control, haploinsufficient (HI) OI patient, dominant negative (DN) OI patient, and primary bone marrow mesenchymal stem cells (hMSC) are shown. ( b ) Relative mRNA expression of chondrocyte markers ( ACAN , SOX9 , COL10A1 , and COL2A1 ) was compared in non-differentiated and differentiated iMSCs (healthy controls (C1, C2, C3), OI patients (P1, P2 and P3)), and hMSC. Relative gene expression was normalized against TBP . Standard deviation (SD) is shown per cell line (n = 3); no SD is present for hMSC (n = 1). Statistical significance is indicated on the graphs ( p -values ≤ 0.05 (*), ≤0.01 (**), ≤0.0001 (****)).

Article Snippet: After 48 h, the chondrogenic induction was started with the chondrogenic differentiation medium (PromoCell, Heidelberg, Germany, C-28012) for three weeks.

Techniques: Biomarker Discovery, Staining, Control, Dominant Negative Mutation, Expressing, Gene Expression, Standard Deviation

Journal: Materials Today Bio

Article Title: Bi-lineage inducible and immunoregulatory electrospun fibers scaffolds for synchronous regeneration of tendon-to-bone interface

doi: 10.1016/j.mtbio.2023.100749

Figure Lengend Snippet: Effect of electrospun fibers scaffolds on chondrogenesis in vitro . (a) Alcain Blue and collagen Ⅱ IHC staining of pellets. (b) Volumes of pellets cultured with different extract induction media. (c) Bern scores for cartilage pellets. (d) Immunofluorescent staining of collagen Ⅱ and Sox9 in BMSCs after 21 days of chondrogenic induction with extracts of electrospun fibers scaffolds. (e)–(g) RT-qPCR measurement of the expression of Col2a1 , Aggrecan , and Sox9 in BMSCs after 21 days of chondrogenic induction with extracts of electrospun fibers scaffolds. (h) Western blotting showing expression of collagen Ⅱ, aggrecan, and Sox9 proteins. (i)–(k) Semiquantitative protein expression levels of collagen Ⅱ, aggrecan, and Sox9. (*p < 0.05, **p < 0.01, and ***p < 0.001).

Article Snippet: Osteogenic and chondrogenic differentiation induction mediums were purchased from Cyagen (Shanghai, China).

Techniques: In Vitro, Immunohistochemistry, Cell Culture, Staining, Quantitative RT-PCR, Expressing, Western Blot