Journal: Frontiers in Veterinary Science

Article Title: Characterisation of canine CD34+/CD45 diminished cells by colony-forming unit assay and transcriptome analysis

doi: 10.3389/fvets.2022.936623

Figure Lengend Snippet: Typical plot profiles of canine haematopoietic stem and progenitor cells (HSPCs). Canine bone marrow-derived mononuclear cells (cBM-MNCs) were stained with 7-aminoactinomycin D (7-AAD), which is a viability dye, phycoerythrin (PE)-labelled anti-canine CD34 monoclonal antibody (mAb), and fluorescein isothiocyanate (FITC)-labelled anti-canine CD45 mAb. (A) Whole viable cells, (B) CD34+ cells, and (C) CD34+/CD45 diminished (CD45dim) cells as analysed by flow cytometry.

Article Snippet: Phycoerythrin-conjugated mouse anti-canine CD34 (clone 1H6, BD Bioscience, diluted 1:20) and fluorescein-isothiocyanate-conjugated rat anti-canine CD45 (clone YKIX716.13, Bio-Rad, Hercules, CA, diluted 1:20) monoclonal antibodies were used for flow cytometry analysis.

Techniques: Derivative Assay, Staining, Flow Cytometry

Journal: Frontiers in Veterinary Science

Article Title: Characterisation of canine CD34+/CD45 diminished cells by colony-forming unit assay and transcriptome analysis

doi: 10.3389/fvets.2022.936623

Figure Lengend Snippet: Haematopoietic colony formation by canine haematopoietic stem and progenitor cells (HSPCs). (A) Microscopic observation of canine haematopoietic colonies (Scale bars: 200 μm). (B) Haematopoietic colonies for every 1,000 cells, as counted for whole viable cells, CD34+ cells, and CD34+/CD45diminished (CD45 dim ) cells. Experiments were performed four times independently. Results are presented as mean ± standard error of the mean (SEM). Statistical significance of total colony counts was assessed by one-way analysis of variance (ANOVA). * p < 0.05, *** p < 0.001.

Article Snippet: Phycoerythrin-conjugated mouse anti-canine CD34 (clone 1H6, BD Bioscience, diluted 1:20) and fluorescein-isothiocyanate-conjugated rat anti-canine CD45 (clone YKIX716.13, Bio-Rad, Hercules, CA, diluted 1:20) monoclonal antibodies were used for flow cytometry analysis.

Techniques:

Journal: Frontiers in Veterinary Science

Article Title: Characterisation of canine CD34+/CD45 diminished cells by colony-forming unit assay and transcriptome analysis

doi: 10.3389/fvets.2022.936623

Figure Lengend Snippet: Global gene expression profiling of canine haematopoietic stem and progenitor cells (HSPCs). (A) Principal component analysis of the global gene expression of canine HSPCs. Gene expression is demonstrated by dots and circles of red (whole viable cells), green (CD34+ cells), and blue [CD34+/CD45 diminished (CD45dim) cells]. (B) The volcano plot shows the results of differentially expressed gene (DEG) analysis between CD34+/CD45dim cells and CD34+ cells. Statistical significance was set as false discovery rate (FDR) <0.1. (C) Gene ontology (GO) and Kyoto Encyclopaedia Gene and Genomes (KEGG)-based pathway enrichment analysis of the downregulated genes of CD34+/CD45dim cells. The candidate terms of biological processes are illustrated with adjusted p -values < −Log10 16 (top). The candidate terms revealed by KEGG pathway analysis are demonstrated (bottom). (D) The volcano plot shows the results of DEG analysis between CD34+/CD45dim cells and whole viable cells. Statistical significance was set as FDR <0.1.

Article Snippet: Phycoerythrin-conjugated mouse anti-canine CD34 (clone 1H6, BD Bioscience, diluted 1:20) and fluorescein-isothiocyanate-conjugated rat anti-canine CD45 (clone YKIX716.13, Bio-Rad, Hercules, CA, diluted 1:20) monoclonal antibodies were used for flow cytometry analysis.

Techniques: Gene Expression

Journal: American Journal of Translational Research

Article Title: Improvement of endometrial thickness and in vitro fertilization outcomes in patients with Asherman’s refractory endometrium using autologous mesenchymal stem cells from the stromal vascular fraction

doi: 10.62347/uagf1249

Figure Lengend Snippet: Figure 2. Characterization of cultured human adipose-derived mesenchymal stem cells (ADMSC) contained in the stromal vascular fraction of the adipose tissue. A. Representative flow cytometry results of MSC markers: CD44, CD45, CD73, and CD90. B. Representative images of ADMSC differentiated into chondrocytes: ADMSC cultured under normal (left) and micro mass conditions (right) stained with Alcian Blue.

Article Snippet: MSCs were stained with antibodies against human CD73, CD90, CD105, CD44, CD45, CD34 (Cat# ab9375, ab133582, ab254022, Abcam), and viability marker VivaFIX (Cat#1351112, BioRad).

Techniques: Cell Culture, Derivative Assay, Flow Cytometry, Staining

Journal: Science Advances

Article Title: Knockdown of deleterious miRNA in progenitor cell–derived small extracellular vesicles enhances tissue repair in myocardial infarction

doi: 10.1126/sciadv.abo4616

Figure Lengend Snippet: ( A ) Immunohistochemical staining of ischemic hearts to detect CD63 + total macrophages and iNOS + CD63 + M1-type macrophages (CD63 for red and iNOS for green staining). Scale bars, 100 μm. ( B ) Quantification of the total number of CD63 + macrophages. *** P < 0.001 and **** P < 0.0001. ( C ) Quantification of the ratio of iNOS − CD63 + cells (M2 type) to iNOS + CD63 + cells (M1 type) ( n = 8). ** P < 0.01 versus M2 macrophage proportion in the PBS group. ( D ) Blood circulating CD45 − CD90 + cell population (MSC) in total PBMCs from cardiac IR rats. * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: Twenty million PBMCs were incubated with allophycocyanin (APC)–conjugated anti-rat CD45 antibody (Miltenyi Biotec, North Rhine-Westphalia, Germany) and fluorescein isothiocyanate (FITC)–conjugated anti-rat CD90 antibody (Abcam, Cambridge, UK).

Techniques: Immunohistochemical staining, Staining