Journal: Materials Today Bio

Article Title: A bioactive composite scaffold enhances osteochondral repair by using thermosensitive chitosan hydrogel and endothelial lineage cell-derived chondrogenic cell

doi: 10.1016/j.mtbio.2024.101174

Figure Lengend Snippet: EndMT-derived tEPCs acquire an MSC-like phenotype. ( a ) Flow cytometry analysis of CD44, CD90, CD105 (mesenchymal stem cell markers), CD34 (hematopoietic and endothelial cell marker), and CD45 (leukocyte marker) expression in tEPCs. The empty areas show isotype control staining. The red-filled areas represent the expression of specific markers. ( b ) Representative immunofluorescence images of tEPC surface markers. tEPCs stain positive for MSC markers (CD44, CD90, and CD105) and negative for endothelial cell markers (CD31, eNOS, VE-cadherin, and vWF) (20 × magnification; scale bar: 50 μm). ( c , d ) CFU efficiency of EPCs and tEPCs, assessing self-renewal through the rate of colony formation in CFU assays. ( c ) Representative colonies of EPCs and tEPCs in 6-well plates (scale bar: 5 mm). ( d ) Columns illustrate the CFU efficiency. Values are reported as the mean ± standard deviation (SD) of six replicates. *** P < 0.001. ( e ) Multilineage differentiation potential of tEPCs induced to differentiate into ( i ) chondrogenic (10 × magnification; scale bar: 100 μm), ( ii ) osteogenic (10 × magnification; scale bar: 100 μm), or ( iii ) adipogenic (40 × magnification; scale bar: 10 μm) lineages. Abbreviations: CD, cluster of differentiation; CFU, colony-forming unit; EndMT, endothelial-to-mesenchymal transition; eNOS, endothelial nitric oxide synthase; EPCs, endothelial progenitor cells; MSCs, mesenchymal stem cells; tEPC, transdifferentiated EPCs; VE-cadherin, vascular endothelial cadherin; vWF, von Willebrand factor.

Article Snippet: Next, 1 × 10 6 tEPCs were suspended in 500 μL of PBS containing 20 μg/mL of fluorescein isothiocyanate (FITC)-conjugated antibodies against CD44 (Novus, Centennial, CO, USA), CD90 (Bioworld, Louis Park, MN, USA), CD105 (Bioss, Woburn, MA, USA), CD34 (Bioss), and CD45 (Bioss).

Techniques: Derivative Assay, Flow Cytometry, Marker, Expressing, Control, Staining, Immunofluorescence, Standard Deviation

Journal: Cells

Article Title: Bone Marrow Aspirate Concentrate Combined with Ultra-Purified Alginate Bioresorbable Gel Enhances Intervertebral Disc Repair in a Canine Model: A Preclinical Proof-of-Concept Study

doi: 10.3390/cells13110987

Figure Lengend Snippet: Cell sorting data on the separation and measurement of CD44-positive and CD45-negative cells from the specimens. We obtained 12 (20-week-old) male rabbits for the experiments. ( a ) Two-dimensional (2D) dot plot represents various cells including CD44-positive and CD45-negative cells and debris. The P1 gate excluded dead cells and debris, and sorted out live cells. SSC-A: Side scatter-area; FSC-A: Forward scatter-area. ( b ) The dot plot in the Q4 gate shows the CD44 (FITC)-positive and CD45 (Cy5.5)-negative cells and the number of these cells was measured. ( c ) The concentration ratio was calculated using the number of CD44-positive and CD45-negative cells per unit volume in BMAC and unconcentrated BM.

Article Snippet: Immunostaining was performed using CD44 (Anti-CD44 Rat-Mono (Hermes-1) FITC; Novus Biologicals, Cat# NBP2-22530F, Centennial, CO, USA), a positive marker, and CD45 (Anti-CD45 Polyclonal Antibody, Cy5.5 Conjugated; Bioss Inc., Cat# bs-0522R-Cy5.5, Woburn, MA, USA), a negative marker in rabbit stem cells [ , ].

Techniques: FACS, Concentration Assay

Journal: Cells

Article Title: Bone Marrow Aspirate Concentrate Combined with Ultra-Purified Alginate Bioresorbable Gel Enhances Intervertebral Disc Repair in a Canine Model: A Preclinical Proof-of-Concept Study

doi: 10.3390/cells13110987

Figure Lengend Snippet: Concentration ratio in the three commercially available BMAC preparation kits.

Article Snippet: Immunostaining was performed using CD44 (Anti-CD44 Rat-Mono (Hermes-1) FITC; Novus Biologicals, Cat# NBP2-22530F, Centennial, CO, USA), a positive marker, and CD45 (Anti-CD45 Polyclonal Antibody, Cy5.5 Conjugated; Bioss Inc., Cat# bs-0522R-Cy5.5, Woburn, MA, USA), a negative marker in rabbit stem cells [ , ].

Techniques: Concentration Assay

Journal: Infection and Immunity

Article Title: Mycobacterium bovis BCG-Infected Mice Are More Susceptible to Staphylococcal Enterotoxin B-Mediated Toxic Shock than Uninfected Mice despite Reduced In Vitro Splenocyte Responses to Superantigens

doi: 10.1128/iai.70.8.4148-4157.2002

Figure Lengend Snippet: FIG. 5. BCG infectious dose correlates with increased expression of the T-cell activation markers CD44, IL-2R , and IL-2R. Unfrac- tionated splenocytes (5 105 cells/tube) from each type of spleen were stained with FITC-labeled anti-mouse CD44, anti-mouse CD25/IL- 2R , or anti-mouse CD122/IL-2R and PE-labeled anti-mouse CD4 or anti-mouse CD8 . The percentages of CD4 CD44hi, IL-2R hi, or IL-2Rhi CD4 (white bars) or CD8 cells (black bars) in the spleen were determined by flow cytometry. Means standard errors of the means of three experiments are shown. , P 0.05; , P 0.01. Purified CD4 T cells from the spleens of BCG HD-infected mice (day 16) were stained with FITC anti-mouse CD44 and were sorted in an EPICS Elite ESP (Beckman Coulter) into CD44low (naïve) and CD44hi

Article Snippet: For flow cytometry analysis, splenocytes (5 105) were stained with all or some of the following MAbs (all at 1:100 dilutions in R10A): fluorescein isothiocyanate (FITC)-conjugated mouse anti-mouse V 8 TCR (F23.1), FITCconjugated rat anti-mouse CD25 (IL-2 R ; 7D4), FITC-conjugated rat antimouse CD122 (IL-2 R ; TM- 1), FITC-conjugated rat anti-mouse I-Ad/I-Ed (2G9) (all from PharMingen), or FITC-conjugated rat anti-mouse CD44 (IM7.8.1; Cedarlane Laboratories, Hornby, Canada).

Techniques: Expressing, Activation Assay, Staining, Labeling, Cytometry, Infection

Journal: Cancers

Article Title: Ex Vivo Behaviour of Human Bone Tumor Endothelial Cells

doi: 10.3390/cancers5020404

Figure Lengend Snippet: Characterization of TECs. ( A ) Expression of endothelial markers in a representative TEC line and HAEC cells, used as a control detected by FACS analysis. FACS analysis of HAECs, and TEC 1, with CD31 antibody at 1st passage and 3th passage. Expression of CD44 antigen in HAECs and TEC 1. ( B ) Bar graph expressing percentage of positive cells to specific antigens analyzed by FACS. Data are expressed as mean ± SE of 10 different TEC lines grouped by histology and three control cell lines. Each experiment was repeated three times. TECs vs . HAECs ( p < 0.001). IgG was used as negative control. ( C ) Representative example of TECs analyzed with CD146 antibody.

Article Snippet: For cytofluorimetric analysis the primary antibodies used were: anti CD31-FITC (R&D Systems, Inc. Minneapolis, MN, USA), anti VE-cadherin-PE (Santa Cruz Biotechnology, Inc. Milan, Italy), anti CD133-PE (Miltenyi Biotech), anti fusin-PE (Santa Cruz Biotechnology), anti CD14-PE (R&D Systems), anti CD44-FITC (R&D Systems), anti CD13-PE/Cy5 (Chemicon International, Temecula, CA, USA), anti CD34-PE, anti CD105-PE, anti CD45-PE, anti CD90-PE, anti CD131-PE, anti CCR7-PE, anti CD133-PE (Miltenyi Biotech.), anti CD146-PE (Miltenyi Biotech.) and anti CD309-PE (Miltenyi Biotech.).

Techniques: Expressing, Control, Negative Control