Journal: bioRxiv

Article Title: Seminal extracellular vesicles from boar AI doses contain fertility-predictive protein and miRNA cargo and improve sperm physiology

doi: 10.64898/2026.03.16.712050

Figure Lengend Snippet: Cytometry plots and histograms of seminal extracellular vesicles (EVs) characterization. A) Flow cytometry beads of know size (200 nm – 2 µm). B) GFP EVs were used to identify EVs population. C) PBS 1X 0.22 μM filtered. D) Negative control of CellTrace Violet (CTV) staining diluted in PBS. E) High fertility (HF) and reduced fertility (RF) unstained EVs. F) HF CTV-stained EVs. G) RF CTV-stained EVs. H) CTV-stained EVs + human anti CD81-APC antibody (1:50). I) CTV-stained EVs + human anti CD63-PE antibody (1:50). J) CTV-stained EVs + human anti Hsp70-FITC antibody (1:50). K) Lysated EVs with Triton 1X + 0.1% SDS. Photodetectors used: Violet Side Scatter – Height (Violet SSC-H), Blue 525-Area (B525-A), Blue 525-Height (B525-H), Violet 450-Height (V450-H), Blue 585-Height (B585-H) and Red 660-Height (R660-H).

Article Snippet: To immunophenotype the EVs, 25 μL of samples (aproximately 2.0 x 10 9 EVs/mL) were incubated 30 min at 38 °C with conjugated antibodies against specific EVs markers: human anti CD63-PE diluted 1:50 (Ref. 130-118-150, Miltenyi Biotec, Germany), human anti CD81-APC 1:50 (Ref. 130-119-825, Miltenyi Biotec) and human anti Hsp70-FITC 1:50 (Ref. 130-124-700, Miltenyi Biotec).

Techniques: Cytometry, Flow Cytometry, Negative Control, Staining

Journal: bioRxiv

Article Title: Generation of Self-Organising Macrovascular Constructs by Bioprinting human iPSC-Derived Mesodermal Progenitor Cells

doi: 10.64898/2026.03.16.712040

Figure Lengend Snippet: (A) IF staining demonstrates progression of vascular network formation from d3 to d10. At day 3 (A1) , CD31 staining (red) appears sparse and discontinuous, predominantly presenting as dot-like signals, with minimal formation of endothelial cell cords or networks. By day 10 (A3) , elongated CD31 + endothelial cells lining the artificial lumen become evident, accompanied by the emergence of capillary-like cell cords and an interconnected, branching vascular network. (B) Morphology and cellular differentiation at d7. (B1) H&E staining of 10 µm sections reveals an immature vascular morphology characterized by a relatively thin vessel wall surrounded by mesoderm-like tissue. (B2) CD31 + endothelial cells (red) line the artificial lumen (AL), while αSMA + smooth muscle-like cells (green) form an early media-like layer. (B3–B5) Cells expressing hematopoietic progenitor cell markers (CD150, CD44 and CD45) as well as Iba1 + macrophage-like cells are present within the wall of the tubular constructs. (B6) Collagen IV (green) deposition indicates early extracellular matrix and basement membrane–like formation, particularly within the intima-like region of the tube wall. (C) Advanced structural organization at day 14. (C1) Vascular constructs show partial narrowing of the AL (H&E). (C2) IF images reveal CD31 + vascular structures within the former AL that are lined by CD31 + endothelial cells (red) and covered by αSMA + pericyte- or smooth muscle–like cells (green). (C3–C6) At day 14, cells expressing hematopoietic progenitor cell markers (CD150, CD44, CD34), as well as Iba1 + macrophage-like cells are still present within the vascular constructs. Collagen IV is prominently deposited around the CD31 + endothelial lining of mid-sized and microvascular structures.

Article Snippet: The following primary antibodies were used: CD31 (M0823, DAKO, Agilent, Santa Clara, CA, USA), αSMA (ab5694, Abcam, Cambridgeshire, UK), CD34 (130-105-830, Miltenyi Biotech), CD150 (PA5-21123, Invitrogen), CD44 (103002, BioLegend, San Diego, CA, USA), Collagen IV (ab6586, Abcam), CD45 (13-9457-82, Invitrogen) and IBA1 (019-19741, Wako/Fuji Film, Neuss, Germany).

Techniques: Staining, Cell Differentiation, Expressing, Construct, Membrane