Journal: iScience

Article Title: HucMSCs can alleviate abnormal vasculogenesis induced by high glucose through the MAPK signaling pathway

doi: 10.1016/j.isci.2024.111354

Figure Lengend Snippet: Generation of BVOs and exploration of the effects of high-glucose conditions on angiogenesis in BVOs (A) Schematic representation of the process for generating BVOs derived from human ESCs. (B) BVOs displayed CD31 staining as a marker of endothelial cells, PDGFR-β staining as a marker of pericytes, and Col IV staining as a marker of the basement membrane. Moreover, BVOs were able to take up DiI-Ac-LDL. (C) The gene expression levels of Pecam1, VE-Cadherin, Pdgfr, α-Sma, Col 4a1 and Col 4a2 in BVOs were higher than those in ESCs. (D) HE staining visualized the lumen (∗) of BVOs. (E) Inverted phase contrast images of VI-BVOs on day 3 after embedding in the hydrogel: control medium group (BVOs-Control) and high-glucose medium group (BVOs-HG). (F) Compared to those in the BVOs-Control group, the CD 31 expression levels and acLDL uptake function of endothelial cells in the BVOs-HG group were significantly decreased. (G) Compared to those in the BVOs-Control group, the expression of the endothelial cell marker genes Pecam1/VE-cadherin and the pericyte marker genes Pdgfr/α-sma in the BVOs-HG group was significantly downregulated in the BVOs-HG group. For (C) and (G), data are represented as mean ± SD. ( n = 3 replicates per condition from 3 independent experiments, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001 from two-tailed Student’s t test)(Scale bar: A, B, E, F = 100 μm, D (left) = 50 μm, D (right) = 20 μm).

Article Snippet: H9 human ESCs (hESCs; WA09/H9; ATCC, USA) were maintained in mTeSR Plus Basal Medium (Stem Cell Technologies, 100–0276, CA, USA) in Matrigel (Corning, Kennebunk, ME, USA)-coated 6-well culture plates.

Techniques: Derivative Assay, Staining, Marker, Membrane, Gene Expression, Control, Expressing, Two Tailed Test

Journal: iScience

Article Title: HucMSCs can alleviate abnormal vasculogenesis induced by high glucose through the MAPK signaling pathway

doi: 10.1016/j.isci.2024.111354

Figure Lengend Snippet: BVOs induction can be divided into two phases: Vasculogenesis and angiogenesis (A) ESC-EBs are spread throughout OCT4+ and SSEA4+ ESCs. (B) Compared with hucMSC-EBs, MI-BVOs had characteristics of mesoderm cells, both of which contained PDGFRα + and FLK1 + mesodermal cells. (C) VI-BVOs not only express vascular-related biomarkers for endothelial cells (CD31) and pericytes (PDGFRβ) but also have the functional characteristics of endothelial cells to take up Ac-LDL. (D) The cells in VI-BVOs have angiogenic potential. (E) In total, 42.4% of the cells in VI-BVOs were endothelial cells, and 8.84% were pericytes. (F) qPCR revealed that the relevant gene expression levels in endothelial cells and pericytes were significantly greater than those in ESCs. Data are represented as mean ± SD. ( n = 3 replicates per condition from 3 independent experiments, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001 from two-tailed Student’s t test). (G) Inverted phase contrast images showing that VI-BVOs-derived single cells can grow and multiply in planar culture. (H) After 3 days of planar culture, 44.3% of the cells derived from VI-BVOs were endothelial cells, and 30.1% were pericytes. (I) The VI-BVOs-derived single-cell population contained many CD31 + endothelial cells. Scale bar: 100 μm.

Article Snippet: H9 human ESCs (hESCs; WA09/H9; ATCC, USA) were maintained in mTeSR Plus Basal Medium (Stem Cell Technologies, 100–0276, CA, USA) in Matrigel (Corning, Kennebunk, ME, USA)-coated 6-well culture plates.

Techniques: Functional Assay, Gene Expression, Two Tailed Test, Derivative Assay