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

Journal: bioRxiv

Article Title: Gram-positive bacteria secrete RNA aptamers to activate human STING for IL-1β release

doi: 10.1101/2021.07.21.453173

Figure Lengend Snippet: (A-D) THP1 (black), CASP1 -/- (blue), ASC -/- (grey), CASP4 -/- (green), cGAS -/- (white) and STING -/- (pattern), MΦs were infected with SA (MOI 10) and GBS (MOI 20), IL-1β production was measured in cell supernetant. (E) THP1, STING -/- , and cGAS -/- MΦs were infected with SA (MOI 10) and GBS (MOI 20), cleaved IL-1β (P17) and caspase-1 (P20) were detected in cell supernetants (Sup) and pro-IL- 1β, pro-caspase-1 and GAPDH in cell lysate by immunoblotting. (F) STING -/- (pattern), STING -/- expressing STING R232 (Black) MΦs were infected with SA (MOI 10) and GBS (MOI 20), IL-1β production was measured in cell supernetant. (G,H) Human blood derived MΦs were pre-treated with increasing concentrations of STING inhibitor (H-151) (100 µM, 50 µM, 10 µM) for 1 h and then infected with SA (MOI 10) and GBS (MOI 20) respectively. IL-1β production was measured in cell supernetant. (I) IL-1β production in wild type THP1 MΦs following transfection with increasing concentrations of SA RNA (RNA+LF) (5 µg/ml, 2.5 µg/ml, 0.1 µg/ml). (J,K,L) THP1 MΦs were pre-treated with increasing concentrations of cytosolic RNase A (RNase A+LF) (100 ng/ml, 50ng/ml, 10 ng/ml) followed by infection with SA. IL-1β production was measured in cell supernetant. Heat inactivated RNase A (100 ng/ml, 50ng/ml, 10 ng/ml) (K) and DNase I (100 ng/ml, 50ng/ml, 10 ng/ml) (L) were used as a control in similar experimental setup. Data shown are mean ±SD (n=3), representative of at least three independent experiments. Asterisks indicate statistically significant differences (∗p < 0.05, ∗∗p < 0.01 and ∗∗∗p < 0.001).

Article Snippet: Experiments with small molecule inhibitors: THP1 MΦ were pre-treated with MCC950 (5 µM, 10 µM) , Dynasore (150, 100 µM) (Enzo), KCL (60 mM, 45 mM, 75 mM), H-151 (100 µM, 50 µM, 10 µM) (ProbeChem) for 1 h prior to the stimulations.

Techniques: Infection, Western Blot, Expressing, Derivative Assay, Transfection