Journal: NPG Asia Materials

Article Title: Spatiotemporal regulation of endogenous MSCs using a functional injectable hydrogel system for cartilage regeneration

doi: 10.1038/s41427-021-00339-3

Figure Lengend Snippet: Fig. 10 MSC homing in cartilage defects at 7 days after the operation. A Immunofluorescent staining for CD44 and CD90. B, C Quantitative results for CD44- and CD90-positive cells.

Article Snippet: At 1 week postoperatively, samples were assessed by immunofluorescence staining for CD44 (1:100, NBP222530F, Novus) and CD90 (1:100, ab225, Abcam) to identify endogenous MSC recruitment.

Techniques: Staining

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:

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: BCG infectious dose correlates with increased expression of the T-cell activation markers CD44, IL-2Rα, and IL-2Rβ. Unfractionated splenocytes (5 × 105 cells/tube) from each type of spleen were stained with FITC-labeled anti-mouse CD44, anti-mouse CD25/IL-2Rα, or anti-mouse CD122/IL-2Rβ and PE-labeled anti-mouse CD4 or anti-mouse CD8α. The percentages of CD4+ CD44hi, IL-2Rαhi, or IL-2Rβhi 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 (memory/effector) populations. Sorted cells were cocultured at 105 cells/well either with bone marrow-derived dendritic cells (5 × 104 cells/well) in the presence or absence of SEB (10 μg/ml) or with M12 B-lymphoma cells (5 × 104 cells/well) in the presence or absence of BCG-Ag (10 μg/ml). IFN-γ levels were determined as described in Materials and Methods. Mean ± standard deviation of triplicate wells is shown.

Article Snippet: For flow cytometry analysis, splenocytes (5 × 10 5 ) were stained with all or some of the following MAbs (all at 1:100 dilutions in R10A): fluorescein isothiocyanate (FITC)-conjugated mouse anti-mouse Vβ8 TCR (F23.1), FITC-conjugated rat anti-mouse CD25 (IL-2 Rα; 7D4), FITC-conjugated rat anti-mouse CD122 (IL-2 Rβ; TM-β1), FITC-conjugated rat anti-mouse I-A d /I-E d (2G9) (all from PharMingen), or FITC-conjugated rat anti-mouse CD44 (IM7.8.1; Cedarlane Laboratories, Hornby, Canada).

Techniques: Expressing, Activation Assay, Staining, Labeling, Flow Cytometry, Purification, Infection, Derivative Assay, Standard Deviation