Journal: bioRxiv

Article Title: Tissue resident macrophages innately develop in a human iPSC-derived cardiac organoid model

doi: 10.1101/2025.05.20.654824

Figure Lengend Snippet: A) Schematic representation of the cardiac organoid differentiation protocol. B) Semiquantitative RT-PCR detecting mesodermal ( TBXT, KDR, GATA4 ), cardiac ( NKX2 . 5, TBX5, ISL1, TNNT2, DES, RYR2 ), and endothelial ( PECAM1 ) marker gene expression. C) H&E staining of organoid section at day 2. D-D’) Immunofluorescence analyses detecting early mesodermal markers TBXT and VIM. E) Immunofluorescence analyses detecting early mesodermal markers GATA6 and KDR. F) H&E staining showing hollow cavity-like structures in cardiac organoids at day 8. G-G’) Immunofluorescence analyses detecting mesenchymal cells (VIM + ) in close association with cardiomyocytes (TNNT2 + ). H) H&E staining of organoid sections at day 35 of culture. I) Immunofluorescence analyses showing close interaction of fibroblasts (VIM + ) and cardiomyocytes (TNNT2 + ) at day 35 of culture. J) Cardiac organoids progressively gain in size (D0 = 12.2 μm ± 6.85; D7 = 1231 ± 20.31; D25 = 1613 ± 45.43). K) Beating frequency increases over culture time starting from 25 bpm (± 1.9) at day 7 up to 57 bpm (± 10.5) at day 35. Scale in all pictures: 50 µm

Article Snippet: Primary antibodies against PECAM1 (CD31) (DAKO, M0823), Vimentin (Invitrogen, MA5-6409), ACTN1 (Abcam, Ab68167), KDR (Miltenyi, 130-125-988), TBXT (R&D Systems, AF2085), TNNT2 (Life Technologies, MA512960), CD34 (Life Technologies, MA5-32059), AIF1 (WAKO/Fuji Film, 019-19741) and CD45 (Life Technologies, 14-0451-82) were diluted in NBS blocking solution and incubated overnight at 4 °C.

Techniques: Reverse Transcription Polymerase Chain Reaction, Marker, Gene Expression, Staining, Immunofluorescence

Journal: Frontiers in Cardiovascular Medicine

Article Title: A refined protocol for the isolation and monoculture of primary mouse renal peritubular endothelial cells

doi: 10.3389/fcvm.2023.1114726

Figure Lengend Snippet: Reagents and resources list for MRPEC isolation.

Article Snippet: CD309 (VEGFR-2) antibody, APC, REAfinity , Recombinant human IgG1; Human anti-Mouse Monoclonal (REA1116) , 1:50 , Miltenyi Biotec , 130-119-434.

Techniques: Isolation, Immunofluorescence, Staining, Flow Cytometry, Recombinant, Concentration Assay, Control, Sterility, Membrane, Cell Culture, Centrifugation, Polymer, Blocking Assay, Purification, Binding Assay, Clinical Proteomics

Journal: Molecular Medicine Reports

Article Title: Implantation of engineered conduction tissue in the rat heart

doi: 10.3892/mmr.2019.9933

Figure Lengend Snippet: Immunohistochemical staining for the expression of vascular markers. Positive staining for CD-31 is shown at days (A) 20, (B) 40 and (C) 60; positive staining for Factor-VIII is shown at days (D) 20, (E) 40 and (F) 60; and positive staining of VEGFR2 is shown at days (G) 20, (H) 40 and (I) 60. The black arrows indicate the presence of the vascular markers (yellow). CD-31, cluster of differentiation 31; VEGFR2, vascular endothelial growth factor receptor 2.

Article Snippet: Excess solution was discarded and the sections were incubated at 4°C overnight with primary antibodies, including rabbit anti-rat CD-31 (cat. no. BA2966; 1:200), Factor-VIII (cat. no. PB0273; 1:100) and VEGFR2 (cat. no. A00901; 1:500; Wuhan Boster Biological Technology, Ltd., Wuhan, China), followed by treatment with goat anti-rabbit peroxidase-conjugated secondary antibody (cat. no. ab6721; 1:1,000; Abcam) for 1 h at room temperature for visualization with 3-diaminobenzidine tetrahydrochloride.

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: The Journal of Physiological Sciences : JPS

Article Title: Senescent endothelial progenitor cells from dogs with pulmonary arterial hypertension: a before–after self-controlled study

doi: 10.1007/s12576-009-0053-7

Figure Lengend Snippet: CD133/KDR-positive cells were detected in peripheral blood and identified as circulating EPCs by flow cytometry using counting beads tubes. a Gated and counted beads from a fluorescein isothiocyanate (FITC) versus side-scattering corner (SSC) dot plot. Quadrants were set on the basis of isotype-identical controls. b Dual-positive CD133/KDR cells were identified and numerated in the upper right quadrant of the scatter plot from blood of dogs. The representative dot plots are represented. The absolute counts of positive cells per ml blood were calculated using the equation described in “Methods”. c Comparison of circulating EPCs number before and after PAH. d Differentiated EPCs were significantly reduced in PAH but transiently raised at the early stage after DHMC. Data are mean ± SD. *p < 0.05 versus baseline, # p < 0.05 versus 48 h

Article Snippet: Briefly, 50 μl of well-mixed anticoagulated whole blood was incubated with PE-labeled mouse anti-dog KDR antibody (Becton Dickinson) and APC-labeled mouse anti-human CD133 antibody (Miltenyi Biotec, Krohne, Germany) in manufacturer-recommended concentrations just above the stainless steel retainer of the tube for 30 min at 4°C.

Techniques: Flow Cytometry, Comparison