Journal: Biomolecules

Article Title: Perlecan in the Natural and Cell Therapy Repair of Human Adult Articular Cartilage: Can Modifications in This Proteoglycan Be a Novel Therapeutic Approach?

doi: 10.3390/biom11010092

Figure Lengend Snippet: Donor demographics and samples.

Article Snippet: At P2, chondrocytes were seeded into 12-well plates at or in chamber slides (with 8 chambers) 5200 cells/cm 2 , and treated with complete media supplemented with or without 200 ng/mL of recombinant active human heparanase (Bio-Techne, Abingdon, UK; 20 ng of enzyme results in >50% of optical density (OD) reduction as measured by heparan sulphate release from human syndecan-4) for 48 h. The 12-well plate was placed in a Cell-IQ (ChipMan Technologies, Tampere, Finland) live imaging platform to acquire phase contrast images of all wells, every ten minutes, during the 48-h culture.

Techniques:

Journal: Biomolecules

Article Title: Perlecan in the Natural and Cell Therapy Repair of Human Adult Articular Cartilage: Can Modifications in This Proteoglycan Be a Novel Therapeutic Approach?

doi: 10.3390/biom11010092

Figure Lengend Snippet: Morphology and growth kinetics of chondrocytes after treatment with heparanase ( n = 5). ( A ) Phase contrast images were acquired for control and heparanase-treated chondrocytes every 10 min at precise locations for 48 h. The representative images shown are of chondrocytes from donor 20 at t = 0 h and t = 48 h. ( B ) Growth kinetics of chondrocytes were established using a live cell imaging platform and analysis software during the 48 h heparanase treatment period. Individual plots are shown for donors 16 to 20 with mean and SD of cell counts from three FOV from three separate wells. The combined data for all five donors at every time point are also shown (bottom right). FOV = field of view. ** p < 0.05, *** p < 0.01.

Article Snippet: At P2, chondrocytes were seeded into 12-well plates at or in chamber slides (with 8 chambers) 5200 cells/cm 2 , and treated with complete media supplemented with or without 200 ng/mL of recombinant active human heparanase (Bio-Techne, Abingdon, UK; 20 ng of enzyme results in >50% of optical density (OD) reduction as measured by heparan sulphate release from human syndecan-4) for 48 h. The 12-well plate was placed in a Cell-IQ (ChipMan Technologies, Tampere, Finland) live imaging platform to acquire phase contrast images of all wells, every ten minutes, during the 48-h culture.

Techniques: Live Cell Imaging, Software

Journal: Biomolecules

Article Title: Perlecan in the Natural and Cell Therapy Repair of Human Adult Articular Cartilage: Can Modifications in This Proteoglycan Be a Novel Therapeutic Approach?

doi: 10.3390/biom11010092

Figure Lengend Snippet: Flow cytometry analysis of the effects of heparanase treatment on surface markers of chondrocytes ( n = 5). Results are shown as the percentage of positive cells for a particular marker on heparanase-treated chondrocytes and the matching control. Fluorochrome-conjugated antibodies were used to detect ( A ) stem cells markers, ( B ) integrins and ( C ) chondrogenic markers. No significant differences were observed. Matched samples are represented by the same colour dot; donor 16 (black), donor 17 (red), donor 18 (blue), donor 19 (green), and donor 20 (yellow).

Article Snippet: At P2, chondrocytes were seeded into 12-well plates at or in chamber slides (with 8 chambers) 5200 cells/cm 2 , and treated with complete media supplemented with or without 200 ng/mL of recombinant active human heparanase (Bio-Techne, Abingdon, UK; 20 ng of enzyme results in >50% of optical density (OD) reduction as measured by heparan sulphate release from human syndecan-4) for 48 h. The 12-well plate was placed in a Cell-IQ (ChipMan Technologies, Tampere, Finland) live imaging platform to acquire phase contrast images of all wells, every ten minutes, during the 48-h culture.

Techniques: Flow Cytometry, Marker

Journal: Biomolecules

Article Title: Perlecan in the Natural and Cell Therapy Repair of Human Adult Articular Cartilage: Can Modifications in This Proteoglycan Be a Novel Therapeutic Approach?

doi: 10.3390/biom11010092

Figure Lengend Snippet: Analysis of the effects of heparanase treatment on gene expression of chondrocytes in monolayer culture ( n = 5). Results are presented as log-fold change in the expression of the chondrogenic genes SOX9, collagen type II, aggrecan, FGFR3 and hypertrophy genes collagen type X and ALK1 in chondrocytes that were treated with heparanase compared to the untreated controls. Matched samples are represented by the same colour dot; donor 16 (black), donor 17 (red), donor 18 (blue), donor 19 (green), and donor 20 (yellow). Error bars indicate medians and interquartile ranges.

Article Snippet: At P2, chondrocytes were seeded into 12-well plates at or in chamber slides (with 8 chambers) 5200 cells/cm 2 , and treated with complete media supplemented with or without 200 ng/mL of recombinant active human heparanase (Bio-Techne, Abingdon, UK; 20 ng of enzyme results in >50% of optical density (OD) reduction as measured by heparan sulphate release from human syndecan-4) for 48 h. The 12-well plate was placed in a Cell-IQ (ChipMan Technologies, Tampere, Finland) live imaging platform to acquire phase contrast images of all wells, every ten minutes, during the 48-h culture.

Techniques: Expressing

Journal: Biomolecules

Article Title: Perlecan in the Natural and Cell Therapy Repair of Human Adult Articular Cartilage: Can Modifications in This Proteoglycan Be a Novel Therapeutic Approach?

doi: 10.3390/biom11010092

Figure Lengend Snippet: Immunocytochemistry and toluidine blue staining of heparanase-treated chondrocytes (two representative donor examples shown). ( A ) Chondrocytes cultured in chamber slides were treated with heparanase for 48 h and stained for perlecan ( n = 4). Chondrocytes in untreated controls tended to remain in clusters that stained more intensely for perlecan compared to heparinase-treated chondrocytes. ( B ) Toluidine blue staining revealed no noticeable trend. However, cell clusters in both controls and heparanase conditions had stronger staining than individual cells. Scale bars = 50 µm.

Article Snippet: At P2, chondrocytes were seeded into 12-well plates at or in chamber slides (with 8 chambers) 5200 cells/cm 2 , and treated with complete media supplemented with or without 200 ng/mL of recombinant active human heparanase (Bio-Techne, Abingdon, UK; 20 ng of enzyme results in >50% of optical density (OD) reduction as measured by heparan sulphate release from human syndecan-4) for 48 h. The 12-well plate was placed in a Cell-IQ (ChipMan Technologies, Tampere, Finland) live imaging platform to acquire phase contrast images of all wells, every ten minutes, during the 48-h culture.

Techniques: Immunocytochemistry, Staining, Cell Culture

Journal: Journal of Cancer Research and Clinical Oncology

Article Title: Fra-2 overexpression upregulates pro-metastatic cell-adhesion molecules, promotes pulmonary metastasis, and reduces survival in a spontaneous xenograft model of human breast cancer

doi: 10.1007/s00432-021-03812-2

Figure Lengend Snippet: Primer sequences of genes used for qRT-PCR

Article Snippet: Tspan8 (RRID: AB_2678899; HPA-044337; 1:50) (Atlas; Merck, Taufkirchen, Germany) , S1699 (DAKO), 125 °C.

Techniques:

Journal: Journal of Cancer Research and Clinical Oncology

Article Title: Fra-2 overexpression upregulates pro-metastatic cell-adhesion molecules, promotes pulmonary metastasis, and reduces survival in a spontaneous xenograft model of human breast cancer

doi: 10.1007/s00432-021-03812-2

Figure Lengend Snippet: Immunohistochemistry of primary tumours engrafted in scid mice and their spontaneous metastases in the lung tissue. a – d IHC staining of representative paraffin-embedded primary tumour tissue and lung tissue with metastasized cells grown in scid mice showing a higher immunoreactivity of L1-CAM, ICAM-1, CD44, and ITGB4 in the tumour cells. Scale bar: 200 and 50 µm. e Images of paraffin-embedded lung sections of scid mice display an immunoreactivity of TSPAN8 within the metastasized cells. This could indicate that these genes are essential for the tumour cells during the emigration process. Arrows label metastasized cells in the lungs. Scale bar: 100 µm

Article Snippet: Tspan8 (RRID: AB_2678899; HPA-044337; 1:50) (Atlas; Merck, Taufkirchen, Germany) , S1699 (DAKO), 125 °C.

Techniques: Immunohistochemistry

Journal: Journal of Cancer Research and Clinical Oncology

Article Title: Fra-2 overexpression upregulates pro-metastatic cell-adhesion molecules, promotes pulmonary metastasis, and reduces survival in a spontaneous xenograft model of human breast cancer

doi: 10.1007/s00432-021-03812-2

Figure Lengend Snippet: Selection of genes, which were deregulated by Fra-2 overexpression in scid mice

Article Snippet: Tspan8 (RRID: AB_2678899; HPA-044337; 1:50) (Atlas; Merck, Taufkirchen, Germany) , S1699 (DAKO), 125 °C.

Techniques: Selection, Over Expression, Biomarker Discovery, Control

Journal: Journal of Cancer Research and Clinical Oncology

Article Title: Fra-2 overexpression upregulates pro-metastatic cell-adhesion molecules, promotes pulmonary metastasis, and reduces survival in a spontaneous xenograft model of human breast cancer

doi: 10.1007/s00432-021-03812-2

Figure Lengend Snippet:

Article Snippet: Tspan8 (RRID: AB_2678899; HPA-044337; 1:50) (Atlas; Merck, Taufkirchen, Germany) , S1699 (DAKO), 125 °C.

Techniques:

Journal: Journal of Cancer Research and Clinical Oncology

Article Title: Fra-2 overexpression upregulates pro-metastatic cell-adhesion molecules, promotes pulmonary metastasis, and reduces survival in a spontaneous xenograft model of human breast cancer

doi: 10.1007/s00432-021-03812-2

Figure Lengend Snippet: Validation of microarray results by qRT-PCR of selected genes

Article Snippet: Tspan8 (RRID: AB_2678899; HPA-044337; 1:50) (Atlas; Merck, Taufkirchen, Germany) , S1699 (DAKO), 125 °C.

Techniques: Biomarker Discovery, Microarray