Journal: ESC Heart Failure

Article Title: Metformin inhibits proliferation of residing fibroadipogenic progenitor cells from failing human hearts

doi: 10.1093/eschf/xvag001

Figure Lengend Snippet: CD45 − CD31 − CD34 + cells isolated from the human myocardium possess fibro- and adipogenic differentiation potential. For subsequent experiments, mononuclear cells were isolated from the left ventricle of human hearts from heart transplantation recipients. CD45 − CD31 − CD34 + cells were further selected by fluorescence-activated cell sorting for in vitro experimentation. ( A ) PDGFRA is a marker of mesenchymal stem cells (multipotent stem cells) and was exclusively expressed in one cluster with a fibroblast phenotype. CD34 was expressed in the same cluster as PDGFRα , as well as in a large cluster of PECAM1 (CD31) positive cells, which represent endothelial cells. PTPRC (CD45) was exclusively expressed in two clusters representing macrophages and T/NK cells. ( B ) Live CD45 − CD31 − CD34 + cells isolated by fluorescence-activated cell sorting. ( C ) Isolated CD45 − CD31 − CD34 + cells can be grown in cell culture and express PDGFRα assessed by immunofluorescence staining (magnification 10X). ( D and E ) Isolated CD45 − CD31 − CD34 + cells increases COL-1 expression when grown in fibrogenic media, suggesting that they differentiate into fibroblasts. ( F and G ) Isolated CD45 − CD31 − CD34 + cells start to express PLIN-1 when grown in adipogenic media, suggesting that they differentiate into adipocytes. Thus, these cells may represent human cFAPs. The image magnifiation for D-G is 20X

Article Snippet: Anti CD45 , Miltenyi , Cat. no. 130-114-123.

Techniques: Isolation, Transplantation Assay, Fluorescence, FACS, In Vitro, Marker, Cell Culture, Immunofluorescence, Staining, Expressing

Journal: ESC Heart Failure

Article Title: Metformin inhibits proliferation of residing fibroadipogenic progenitor cells from failing human hearts

doi: 10.1093/eschf/xvag001

Figure Lengend Snippet: Metformin in pharmacological doses (0.1 mM) inhibits proliferation of fibrogenic progenitors. Mononuclear non-muscle cells were isolated from the left ventricle of explanted human hearts from heart transplantation recipients. Subsequently, CD45 − CD31 − CD34 + cells were isolated by fluorescence-activated cell sorting for in vitro experimentation. ( A and B ) COL-1 and COL-6 mRNA expression in cFAPs grown in fibrogenic media and incubated with metformin (0.1 mM) and cimetidine (0.1 mM) alone and in combination. ( C ) Quantification of proliferation assay as per cent 5-ethynyl-2'-deoxyuridine (EDU) positive cells. Individual donors ( A–C ) are indicated by the shape of the dots. ( D ) Cell proliferation assays (EDU) and apoptosis assays (TUNEL) performed on cFAPs in cell culture incubated with metformin (0.1 mM) and cimetidine (0.1 mM) alone and in combination (magnification 10X) (* P < .05)

Article Snippet: Anti CD45 , Miltenyi , Cat. no. 130-114-123.

Techniques: Isolation, Transplantation Assay, Fluorescence, FACS, In Vitro, Expressing, Incubation, Proliferation Assay, TUNEL Assay, Cell Culture

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Reconstitution of Circulating Mucosal-Associated Invariant T Cells after Allogeneic Hematopoietic Cell Transplantation: Its Association with the Riboflavin Synthetic Pathway of Gut Microbiota in Cord Blood Transplant Recipients.

doi: 10.4049/jimmunol.1900681

Figure Lengend Snippet: FIGURE 3. Comparison of MR1 tetramer+ MAIT cells and CD161+TCR Va7.2+MAIT cells in long-term survivors after allogeneic HCT. (A) Gating strategies for CD161+TCR Va7.2+MAIT cells and MR1: 5-OP-RU and 6-FP tetramer+ MAIT cells. (B) Correlation of proportion of MR1: 5-OP-RU tetramer+ MAIT cells, CD161+TCR Va7.2+ MAIT cells, CD4+CD161+T cells, CD8+CD161+T cells, and MR-1: 6-FP tetramer+ MAIT cells among CD3+

Article Snippet: To examine the proportion of MAIT cells, PBMCs were stained with allophycocyanin-Cy7-conjugated anti-human CD3 (clone SK7; BioLegend, San Diego, CA), FITC-conjugated anti-human TCR g/d (clone B1; BioLegend), PE-conjugated anti-human CD161 (clone HP-3G10; Tonbo Biosciences, San Diego, CA), BV421-conjugated anti-human TCR Va7.2 (clone 3C10; BioLegend), BV510-conjugated anti-human CD4 (clone RPA-T4; BioLegend), PE-Cy7-conjugated anti-human CD8a (clone RPA-T8; Tonbo Biosciences), allophycocyanin-conjugated antihuman programmed cell death–1 (PD-1) (clone EH12.2H7; BioLegend), and PerCP/Cy5.5-conjugated anti-human TCR Va24-Ja18 (clone 6B11; BioLegend) Abs.

Techniques: Comparison

Journal: Journal of Clinical Laboratory Analysis

Article Title: High‐dimensional single‐cell proteomics analysis reveals the landscape of immune cells and stem‐like cells in renal tumors

doi: 10.1002/jcla.23155

Figure Lengend Snippet: Purified antibodies about the immune cell‐centric panel

Article Snippet: CD161 , 164Dy , HP‐3G10 , Fluidigm , 3164009B.

Techniques: Purification

Journal: European journal of haematology

Article Title: Morphologic, flow cytometric, functional, and molecular analyses of S100B positive lymphocytes, unique cytotoxic lymphocytes containing S100B protein.

doi: 10.1111/ejh.12036

Figure Lengend Snippet: Figure 1 Immunophenotyping of S100B+ lymphocytes by flow cytometry. The peripheral blood lymphocytes (PBL) of a healthy adult were stained for various lymphocyte surface markers and cytoplasmic S100B protein. A three-color flow cytometric analysis of the PBL for S100B/ CD3/CD8 (A), S100B/CD3/CD16 (B), S100B/CD56/CD16 (C), and a two-color analysis for S100B/CD161 (D), S100B/CD244 (E), S100B/CD314 (F), S100B/CD4 (G), S100B/CD25 (H), S100B/CD19 (I), and S100B/HLA-DR (J) were performed. A and B: There are two subtype of S100B+ lympho- cytes, the S100B+ CD3+ CD8+ CD16−subtype (CTL subtype; red dots) and the S100B+ CD3- CD8- CD16+ subtype (NK subtype; blue dots). C: The NK subtype of S100B lymphocytes also express CD56.

Article Snippet: In this study, we attempted to precisely characterize the S100B+ lymphocytes in healthy adult individuals with respect to the proportion, immunophenotypes, responses for Table 1 List of the antibodies used in this study Antibodies Clone Sources Polyclonal Rabbit Anti-Human S100B A5110 Dako Cytomation (Glostrup, Denmark) PE-Conjugated Monoclonal Mouse Anti-Human CD3 UCHT1 BD Pharmingen (Franklin Lakes, NJ, USA) PE-Conjugated Monoclonal Mouse Anti-Human CD4 RPA-T4 BD Pharmingen PE-Conjugated Monoclonal Mouse Anti-Human CD244 2–69 BD Pharmingen PE-Conjugated Monoclonal Mouse Anti-Human HLA-DR L243 BD Pharmingen RPE-Conjugated Monoclonal Mouse Anti-Human CD8 DK25 Dako Cytomation RPE-Conjugated Monoclonal Mouse Anti-Human CD16 DJ130c Dako Cytomation RPE-Conjugated Monoclonal Mouse Anti-Human CD25 ACT-1 Dako Cytomation RPE-Conjugated Monoclonal Mouse Anti-Human CD19 HD37 Dako Cytomation Phycoerythin (PE) anti-human CD56 CMSSB eBioscience (San Diego, CA, USA) Phycoerythin (PE) anti-human CD314/ NKG2D 1D11 eBioscience Anti-Human CD19 APC HIB19 eBioscience Phycoerythin (PE) anti-human αβTCR IP26 eBioscience Phycoerythin (PE) anti-human cdTCR B1.1 eBioscience PE-Conjugated Monoclonal Mouse Anti-Human CD161 191B8 Beckman Coulter (San Jose, CA, USA) RPE-Conjugated Monoclonal Mouse Anti-Human TCR Vα24 C15 AbD Serotec (Kidlington, UK) FITC Goat Anti-Rabbit IgG(H+L) Conjugated Invitrogen (Carlsbad, CA, USA) Streptavidin PerCP BD Pharmingen Biotin Mouse Anti-Human CD3 HIT3a BD Pharmingen Biotin Mouse Anti-Human CD8 RPA-T8 BD Pharmingen Biotin Mouse Anti-Human CD16 3G8 BD Pharmingen 100 © 2012 John Wiley & Sons A/S T-cells mitogen, the morphological response to NK-sensitive or -resistant target cells, and their S100B mRNA expression.

Techniques: Cytometry, Staining