a primary antibody against collagen type ii  (Thermo Fisher)

Journal: Cells

Article Title: Therapeutic Potential of Olfactory Ensheathing Cells and Adipose-Derived Stem Cells in Osteoarthritis: Insights from Preclinical Studies

doi: 10.3390/cells13151250

Figure Lengend Snippet: Adipose stem cell (ASC) characterization. ( A , B ) morphology of ADSC. Scale bar = 100 μm. ( C ) Flowcytometry of ASC. Positive for CD44, CD73, CD90, CD105, and HLA-ABC and negative for CD34, CD45, and HLA-DR. ( D – F ) qRT-PCR showed the gene expressions of adipogenesis ( FABP4 , PPARγ ) ( D ), osteogenesis ( ALPL , RUNX2 ) ( E ), and chondrogenesis (aggrecan and COL2A1 ) ( F ). ( G ) Oil red staining of differentiated cells after adipogenesis. ( H ) Quantification of Oil Red at OD 510. ( I ) Alizarin Red staining of differentiated cells after osteogenesis. ( J ) Quantification of Alizarin Red at OD552. ( K – O ) Characterization of differentiated cell after chondrogenesis. Pellet morphology ( K ), H & E ( L ), Safranin O ( M ), aggrecan ( N ), and type II collagen ( O ). ** p < 0.01, *** p < 0.001. Scale bar = 100 μm.

Article Snippet: Subsequently, the sections were blocked with Ultra V block (Thermo Scientific) for 10 min and then incubated with primary antibodies against type II collagen and aggrecan (1:200, GeneTex, Irvine, CA, USA) at 37 °C for 4 h. The secondary antibodies used were biotin-labeled goat anti-rabbit immunoglobulin (Dako, Carpinteria, CA, USA) and horseradish peroxidase-conjugated streptavidin (Biocare Medical), which were incubated for 30 min.

Techniques: Quantitative RT-PCR, Staining

Journal: Cells

Article Title: Therapeutic Potential of Olfactory Ensheathing Cells and Adipose-Derived Stem Cells in Osteoarthritis: Insights from Preclinical Studies

doi: 10.3390/cells13151250

Figure Lengend Snippet: Immunohistochemistry of type II collagen and aggrecan of the mouse osteoarthritis model after 28 days of experiments. ( A ) A representative image of type II collagen is illustrated from the normal knee ( n = 4), osteoarthritis (OA) ( n = 4), 5 × 10 5 olfactory ensheathing cells (OECs), 1 × 10 5 OECs, and 1 × 10 5 adipose-derived stem cells (ADSCs) ( n = 4 in each group). ( B ) Quantitative analysis of positive staining cells in 50 cells in five fields. ( C ) A representative image of aggrecan is illustrated from the normal knee ( n = 4), OA ( n = 4), 5 × 10 5 OECs, 1 × 10 5 OECs, and 1 × 10 5 ADSCs ( n = 4 in each group). ( D ) Quantitative analysis of positive staining cells in 50 cells in five fields. * p < 0.05, ** p < 0.01.

Article Snippet: Subsequently, the sections were blocked with Ultra V block (Thermo Scientific) for 10 min and then incubated with primary antibodies against type II collagen and aggrecan (1:200, GeneTex, Irvine, CA, USA) at 37 °C for 4 h. The secondary antibodies used were biotin-labeled goat anti-rabbit immunoglobulin (Dako, Carpinteria, CA, USA) and horseradish peroxidase-conjugated streptavidin (Biocare Medical), which were incubated for 30 min.

Techniques: Immunohistochemistry, Derivative Assay, Staining