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

Journal: bioRxiv

Article Title: Rostrocaudal patterning and neural crest differentiation of human pre-neural spinal cord progenitors in vitro

doi: 10.1101/2020.06.16.155564

Figure Lengend Snippet: A, B) Optimal CHIR concentration (3 μM) was optimised in WA01 (H1) hESCs. (A) Representative immunostaining of NMP markers SOX2 (red) and CDX2 (grey) and TBXT (magenta) at 36h after following treatment scheme with 3 μM CHIR and (B) quantification markers over a range of CHIR concentrations between 1-10 μM. Scale bars, 100μm. C,D) Transcriptional analysis (RT-qPCR) of NMP markers TBXT, SOX2 and CDX2 (C) and selected HOX genes (D) up to passage 10. Expression levels are normalised to the reference gene PBDG . Error bars show SD, (n=3 technical replicates). E, F) Optimal CHIR concentration (4 μM) was optimised in AICS ZO1-mEGFP (AICS-0024) iPSCs hESCs. (E) Representative immunostaining of NMP markers SOX2 (red) and CDX2 (grey) and TBXT (magenta) at 36h after following treatment scheme with 3 μM CHIR and (F) quantification markers over a range of CHIR concentrations between 1-10 μM. Scale bars, 100μm. G,H) Transcriptional analysis (RT-qPCR) of NMP markers TBXT, SOX2 and CDX2 (G) and selected HOX genes (H) up to passage 10. Expression levels are normalised to the reference gene PBDG . Error bars show SD, (n=3 technical replicates).

Article Snippet: Human ESCs (WA09 and WA01, WiCell) and human iPSCs (AICS-23, Allen Institute) were maintained in feeder-free cultures, plated on Corning Matrigel Growth Factor Reduced (GFR) Basement Membrane Matrix (Corning Incorporated, 354230) and grown in mTESR1 (STEMCELL technologies, 85850).

Techniques: Concentration Assay, Immunostaining, Quantitative RT-PCR, Expressing