primary human aortic ecs  (PromoCell)

Journal: bioRxiv

Article Title: A Long-lived Avatar for Modeling Age-Related Vascular Disease

doi: 10.64898/2026.04.29.721776

Figure Lengend Snippet: A , Representative images of iPSC-ECs in low serum (3%) and treated with VEGF, SB, or VEGF+SB (VSL) at the indicated time points reveals that VSL further improves EC morphology. The above treatments were performed using EC medium (Lonza, EGM-2MV) with 3% FBS. Ctrl* = EC medium with 3% FBS; VEGF = VEGF (10 ng/mL) added to EC medium with 3% FBS; SB = SB (SB 431542, 10 µM) added to EC medium with 3% FBS; VEGF + SB = VEGF (10 ng/mL) and SB (SB 431542, 10 µM) added to EC medium with 3% FBS, referred to as VSL. (scale bar: 100 µm) B , Quantification of cell circularity from ( A ) reveals that VSL reduces cell circularity. C , Quantification of cell area from ( A ) reveals that VSL reduces cell area. D , UMAP analysis of iPSC-ECs treated with different viability factors or combinatorial treatments of viability factors at day 40 shows that VSL restores the transcriptional profile of HAECs at day 40 to that at day 0 (n=3). The prophylactic antibiotic (Lonza Walkersville MycoZap, 0.2%; AB) was used to prevent mycoplasma contamination. D0_Ctrl = iPSC-ECs at day 0; D40_Ctrl = iPSC-ECs treated with 3%FBS EC medium (Lonza, EGM-2MV) for 40 days; D40_VEGF = iPSC-ECs treated with the 3%FBS EC medium with the addition of VEGF (10 ng/mL) for 40 days; D40_AB = iPSC-ECs treated with 3%FBS EC medium + prophylactic antibiotic (Lonza Walkersville MycoZap, 0.2%; AB) for 40 days; D40_VSL = iPSC-ECs treated with VSL for 40 days; D40_VSL+cAMP = iPSC-ECs treated with VSL and cAMP (10 µM) for 40 days; D40_VSL+cGMP = iPSC-ECs treated with VSL and cGMP (10 µM) for 40 days; D40_VSL+Camp + cGMP = iPSC-ECs treated with VSL, cAMP (10 µM), and cGMP (10 µM) for 40 days; D40_VSL + cAMP + cGMP + AB = iPSC-ECs treated with VSL, cAMP (10 µM), cGMP (10 µM) and AB (Lonza Walkersville MycoZap, 0.2%;) for 40 days. E , Representative images of β-gal staining for iPSC-ECs treated with or without VSL at day 40 are shown (scale bar: 200 µm). F , Quantification of (E) shows that VSL reduces the percentage of β-gal positive cells (n-=3). Each dot represents one single field. G , The expression of sICAM-1 and sVCAM-1 in culture media from iPSC-ECs treated with VEGF alone or VSL at day 0 or day 20 detected by ELISA assay shows that VSL effectively suppresses inflammatory cytokines (n=3). D0_Ctrl = iPSC-ECs at day 0; D20_ Ctrl = iPSC-ECs treated with standard EC medium (Lonza, EGM-2MV) for 20 days; D20_VEGF = iPSC-ECs treated with standard EC medium and addition of VEGF (10 ng/mL) for 20 days; D20_VSL = iPSC-ECs treated with VSL for 20 days. Each dot represents one technical repeat. H , Heatmap analysis of EC marker genes and fibroblast marker genes expressed in iPSC-ECs treated with or without VSL at day 60 reveals that VSL preserves EC identity during long-term culture. Each group includes triplicate in this analysis. Ctrl_D60 = iPSC-ECs treated with 3%FBS EC medium (Lonza, EGM-2MV) for 60 days. VSL_D60 = iPSC-ECs treated with VSL. Data between two groups were analyzed by Student’s t-test. Data between multiple groups are analyzed by one-way ANOVA. Results are considered statistically significant with P<0.05(*), P<0.01(**), P<0.001(***), and P<0.0001(****).

Article Snippet: HAECs were purchased from ATCC (Cat# PCS-100-011). iPSC-ECs were generated using established methods in our laboratory .

Techniques: Staining, Expressing, Enzyme-linked Immunosorbent Assay, Marker

Journal: bioRxiv

Article Title: A Long-lived Avatar for Modeling Age-Related Vascular Disease

doi: 10.64898/2026.04.29.721776

Figure Lengend Snippet: A , mVSL+ maintains co-culture of human iPSC-ECs and iPSC-VSMCs for over 180 days, with endothelial cells aligning with the lumen direction throughout the 180-day culture (scale bar: 40 µm). B , Quantification of ( A ) shows that mVSL+ preserves lumen surface integrity throughout 180-day culture. C, Immunofluorescence staining of VE-Cadherin and α-SMA in the avatars at the indicated time points (scale bar: 40 µm). D , mVSL+ maintains the expression of sICAM-1 and sVCAM-1 in the vascular avatars at a relatively low level compared to day 5 throughout the 180-day culture. (n=3). E , Bio-plex analysis shows that mVSL+ maintains the vascular avatars at a low inflammatory state compared to day 5 throughout the 180-day culture . F , The duration of prior 3D models of vascular lumens using monoculture (in red) or co-culture (in blue). Each dot represents the duration noted in one paper. 17 papers are cited in total. The large red dot indicates our monoculture duration, and the large blue dot indicates our co-culture duration.

Article Snippet: HAECs were purchased from ATCC (Cat# PCS-100-011). iPSC-ECs were generated using established methods in our laboratory .

Techniques: Co-Culture Assay, Immunofluorescence, Staining, Expressing

Journal: Redox Biology

Article Title: Big data analytics for MerTK genomics reveals its double-edged sword functions in human diseases

doi: 10.1016/j.redox.2024.103061

Figure Lengend Snippet: Proteomics in HAECs with MerTK gene knockout or control. ( A ) MerTK expression in HAECs incubated with apoptotic Jurkat cells for 1 h. ( B ) Immunochemical staining for MerTK expression in the aortic arch from WT mice. ( C ) Efferocytosis of apoptotic Jurkat cells by HAECs after 1 h of co-incubation. P < Apoptotic Jurkat cells were labeled with green PKH67 (Sigma) and HAECs were labeled with red PKH26 (Sigma). Green cells are apoptotic Jurkat cells that were not engulfed by HAECs. Green/red small round cells are apoptotic Jurkat cells that were engulfed by HAECs. Large red cells are HAECs. (D) Volcano plot illustration in MerTK KO vs. control. Relative protein abundance (log2) plotted against significance level (-log10 P-value), showing significantly (p < 0.05) downregulated (blue), upregulated (red) or non-differentially expressed proteins (grey). (E) Graphic summarization for pathways in MerTK KO vs. control. (F) MerTK KO activates apoptosis signaling. (G) Canonical pathway analysis in MerTK KO vs. control. Color depends on z-score. Blue signifies negative value; orange signifies positive value; and grey signifies no activity pattern. Size is proportional to the number of genes that overlap the pathway. (H) Machine learning analysis for activated or inhibited disease pathways. (I–K) IPA prediction shows that MerTK KO activates premature aging, kidney failure and heart failure. Proteomics data were analyzed by IPA. Data were analyzed with GraphPad Prism 9.4.1 and shown as the mean ± SD (n = 3–5). P < 0.05 was considered statistically significant. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Primary human aortic ECs (HAECs) and the human Jurkat cell line were purchased from ATCC (Manassas, VA, USA).

Techniques: Gene Knockout, Control, Expressing, Incubation, Staining, Labeling, Quantitative Proteomics, Activity Assay