primary human dermal microvascular ecs  (PromoCell)

Journal: bioRxiv

Article Title: Mitochondrial Dysfunction in Endothelial Cells Drives Greater Vascular Impairment in Females with Diabetes-Associated Peripheral Artery Disease

doi: 10.1101/2025.09.22.677648

Figure Lengend Snippet: (A) Schematic illustrating site of tissue collection from below- and above knee amputations. (B) Myography showing increased endothelial dysfunction in female vs male patient arteries. Left , endothelial-dependent relaxation in response to acetylcholine (ACh). Right , No difference in vascular smooth muscle cell (VSMC), sodium nitroprusside (SNP)- mediated relaxation (n=3-5). (C) Left, representative image of microvessels in tibialis anterior (CD31 + SMA + , yellow arrows). Laminin (myocytes, white), CD31 (ECs, red) and SMA (pericytes, green); scale bar 50 μm. Right , quantification (n=5/sex). (D) Myocyte area is unaltered with sex (n=5). (E) NADPH oxidase ( Nox ) mRNA marker expression in muscle measured by qPCR, normalized to β -actin (n=5). (F) 8-isoprostane levels in plasma remain unchanged with sex (n=5). Results are mean±SEM; two-way ANOVA, Student’s t -test or Mann–Whitney U -test; * P <0.05, ** P <0.01, *** P <0.001.

Article Snippet: Primary human male and female coronary artery ECs were purchased from American Type Culture Collection.

Techniques: Marker, Expressing, Clinical Proteomics, MANN-WHITNEY

Journal: bioRxiv

Article Title: Mitochondrial Dysfunction in Endothelial Cells Drives Greater Vascular Impairment in Females with Diabetes-Associated Peripheral Artery Disease

doi: 10.1101/2025.09.22.677648

Figure Lengend Snippet: (A) Schematic to show vessel collection for myography. (B) Myography showing increased endothelial dysfunction in female vs male arteries. Left , endothelial-dependent relaxation in response to acetylcholine (ACh). Right , sodium nitroprusside (SNP)-mediated relaxation (n=5-7). (C) Myography in non-injured vessels (N=5-7). (D) Plasma nitrite/nitrate levels (n=4-6). Laser Doppler imaging showing reduced blood perfusion over time with diabetes in (E) male and (F) female mice. Left, representative image of blood flow at 14 d. Right, quantification. (n=9-11). (G) Laser doppler perfusion index at 14 days (n=9-11). (H) Microvessel number (CD31+SMA+, <50 µm in diameter) in gastrocnemius tissues normalized to the number of myocytes and control non-ischemic limbs (n=9-11). (I) Tubule formation of male and female murine ECs shows an altered phenotype. (J) Tubulogenesis is reduced in female ECs under diabetic conditions. Left, Wimasis platform for vessel coverage and networks. Right , quantification (n=5/group). Results are mean±SEM; two-way ANOVA, Student’s t -test or Mann–Whitney U -test; * P <0.05, ** P <0.01, *** P <0.001 and **** P <0.0001.

Article Snippet: Primary human male and female coronary artery ECs were purchased from American Type Culture Collection.

Techniques: Clinical Proteomics, Imaging, Control, MANN-WHITNEY

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: TGF-β1 Promotes Angiogenesis via Endothelial-to-Mesenchymal Transition in Infantile Hemangioma

doi: 10.1161/ATVBAHA.125.322793

Figure Lengend Snippet: Changes in the expression of endothelial and mesenchymal markers after TGF-β1 (transforming growth factor β1) treatment in hemangioma-derived endothelial cells (HemECs) and human umbilical vein endothelial cells (HUVECs). A , Western blot analysis showing the expression of TGF-β1 in HemECs and HUVECs (n=4 biological replicates). The unpaired 2-tailed Student t tests were used. P =0.001, Cohen d =−7.50 (significant, extremely large effect). B , Immunofluorescence images showing the expression of endothelial and mesenchymal markers before and after TGF-β1 treatment in HemECs and HUVECs (n=4 biological replicates). Scale bar, 50 µm. C , Immunofluorescence analysis showing the mean fluorescence intensity of endothelial and mesenchymal markers before and after TGF-β1 treatment (n=4 biological replicates). The unpaired 2-tailed Student t tests were used. For marker expression in HemECs (TGF-β1 vs untreated), CD31: P =0.001, Cohen d =−7.14 (significant, extremely large effect). VE-cadherin (vascular endothelial cadherin): P =0.030, Cohen d =−2.93 (significant, very large effect). α-SMA (α-smooth muscle actin): P <0.001, Cohen d =13.55 (significant, extremely large effect). COL1A1 (collagen type I alpha 1 chain): P <0.001, Cohen d =11.58 (significant, extremely large effect). For marker expression in HUVECs (TGF-β1 vs untreated), CD31: P =0.72, Cohen d =−1.14 (not significant, large effect). VE-cadherin: P =0.21, Cohen d =1.65 (not significant, very large effect). α-SMA: P =0.92, Cohen d =−0.08 (not significant, negligible effect). COL1A1: P =0.93, Cohen d =−0.11 (not significant, negligible effect).

Article Snippet: HemEC isolation from proliferating IHs was performed as described previously., Primary human umbilical vein ECs (HUVECs) were obtained from the American Type Culture Collection (United States).

Techniques: Expressing, Derivative Assay, Western Blot, Immunofluorescence, Fluorescence, Marker

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: TGF-β1 Promotes Angiogenesis via Endothelial-to-Mesenchymal Transition in Infantile Hemangioma

doi: 10.1161/ATVBAHA.125.322793

Figure Lengend Snippet: TGF-β1 OE (TGF-β1 [transforming growth factor β1] overexpression) promotes hemangioma-derived endothelial cell (HemEC) migration, invasion, and angiogenesis. A , Transwell migration assay results showing the migration ability of HemECs and human umbilical vein endothelial cells (HUVECs) after TGF-β1 OE (n=4 biological replicates). Scale bar, 100 µm. The unpaired 2-tailed Student t tests were used. HUVECs: TGF-β1 OE vs control: P =0.96, Cohen d =0.44 (not significant, small to medium effect). HemECs: TGF-β1 OE vs control: P <0.001, Cohen d =8.85 (significant, extremely large effect). B , Transwell invasion assay results showing the invasion ability of HemECs and HUVECs after TGF-β1 OE (n=4 biological replicates). Scale bar, 100 µm. The unpaired 2-tailed Student t tests were used. HUVECs: P =0.62, Cohen d =−0.58 (not significant, medium effect). HemECs: P <0.001, Cohen d =9.29 (significant, extremely large effect). C , Tube formation assay showing the angiogenic ability of HemECs in vitro after TGF-β1 OE (n=4 biological replicates). Formed vessels are marked with yellow arrows. Scale bar, 100 µm. The unpaired 2-tailed Student t tests were used. For total tube length (TGF-β1 OE vs control), HUVECs: P =0.42, Cohen d =−0.79 (not significant, medium effect). HemECs: P <0.001, Cohen d =8.37 (significant, extremely large effect). For the number of branch sites (Control vs TGF-β1 OE ), HUVECs: P =0.60, Cohen d =−0.47 (not significant, medium effect). HemECs: P =0.017, Cohen d =3.19 (significant, extremely large effect). D , Vessel formation results showing the angiogenic ability of HemECs in vivo after TGF-β1 OE using 4 mice in each group (n=4). Scale bar, 50 µm. The unpaired 2-tailed Student t tests were used. TGF-β1 OE vs control: P =0.001, Cohen d =7.25 (significant, extremely large effect).

Article Snippet: HemEC isolation from proliferating IHs was performed as described previously., Primary human umbilical vein ECs (HUVECs) were obtained from the American Type Culture Collection (United States).

Techniques: Over Expression, Derivative Assay, Migration, Transwell Migration Assay, Control, Transwell Invasion Assay, Tube Formation Assay, In Vitro, In Vivo

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: TGF-β1 Promotes Angiogenesis via Endothelial-to-Mesenchymal Transition in Infantile Hemangioma

doi: 10.1161/ATVBAHA.125.322793

Figure Lengend Snippet: Changes in lipid metabolism. A , Oil Red O (ORO) staining showing the number of intracellular lipid droplets (LDs) in hemangioma-derived endothelial cells (HemECs) compared with that in human umbilical vein endothelial cells (HUVECs), with the etomoxir treatment group serving as a positive control (n=4 biological replicates). Scale bar, 20 µm. The Dunnett multiple comparisons test was used. HUVECs: TGF-β1 OE (transforming growth factor β1 overexpression) vs control: P =0.16, Cohen d =1.20 (not significant, large effect). Etomoxir vs control: P <0.001, Cohen d =10.54 (significant, extremely large effect). HemECs: TGF-β1 OE vs control: P =0.001, Cohen d =13.38 (significant, extremely large effect). Etomoxir vs control: P <0.001, Cohen d =24.63 (significant, extremely large effect). B , Western blot analysis showing that CPT1A (carnitine palmitoyltransferase 1A) protein expression decreased in HemECs (n=4 biological replicates). The unpaired 2-tailed Student t test was used. HemEC TGF-β1 OE vs control: P =0.001, Cohen d =−5.84 (significant, extremely large effect). C , Targeted metabolic analysis showing the top 30 differential metabolites (DMs) after TGF-β1 (transforming growth factor β1) treatment in HUVECs (n=6 biological replicates). D , Targeted metabolic analysis showing the top 30 differential metabolites (DMs) after TGF-β1 treatment in HemECs (n=6 biological replicates). E , Quantitative analysis of L-palmitoylcarnitine, with the etomoxir treatment group serving as a positive control (n=6 biological replicates). One-way ANOVA followed by the Dunnett multiple comparisons test was used to compare the TGF-β1 and etomoxir groups to the control group. HUVECs: TGF-β1 vs control: P =0.18, Cohen d =−0.83 (not significant, medium effect). Etomoxir vs control: P <0.001, Cohen d =−2.24 (significant, extremely large effect). HemECs: TGF-β1 vs control: P =0.003, Cohen d =−3.06 (significant, extremely large effect). Etomoxir vs control: P <0.001, Cohen d =−4.76 (significant, extremely large effect). F , Changes in the content of long-chain fatty acids (FAs; chain lengths C 14 –C 18 ; n=4 biological replicates). The paired 2-tailed Student t test with the Welch correction was performed separately for each FA chain length (C14, C16, and C18) to compare the TGF-β1–treated and control groups within HemECs and HUVECs. HemECs: C14: P =0.012, Cohen d =2.50 (significant, extremely large effect). C16: P =0.039, Cohen d =1.73 (significant, extremely large effect). C18: P =0.027, Cohen d =1.89 (significant, extremely large effect). HUVECs: C14: P =0.09, Cohen d =−1.37 (not significant, very large effect). C16: P =0.048, Cohen d =−1.60 (significant, large effect). C18: P =0.049, Cohen d =−1.64 (significant, extremely large effect). G , Palmitate-conjugated BSA (Palm-BSA) stimulated oxygen consumption rate (OCR) in HUVECs (n=4 biological replicates). A paired 2-tailed Student t test was performed. Control: P =0.009, Cohen d =4.11 (significant, extremely large effect). TGF-β1: P =0.010, Cohen d =0.55 (significant, medium effect). Etomoxir: P =0.84, Cohen d = –0.08 (not significant, negligible effect). TGF-β1+etomoxir: P =0.22, Cohen d =1.24 (not significant, large to very large effect). BSA: TGF-β1 vs control: P =0.36, Cohen d =−0.70 (not significant, medium effect). Palm-BSA: TGF-β1 vs control: P =0.08, Cohen d =−4.43 (not significant, extremely large effect). Etomoxir vs control: P <0.001, Cohen d =−4.83 (significant, extremely large effect). TGF-β1+etomoxir vs control: P =0.003, Cohen d =−3.30 (significant, extremely large effect). H , Palm-BSA failed to stimulate the OCR, thus inhibiting the fatty acid oxidation (FAO) effect of TGF-β1 or etomoxir treatment in HemECs (n=4 biological replicates). A paired 2-tailed Student t test was performed. Control: P =0.015, Cohen d =1.73 (significant, extremely large effect). TGF-β1: P =0.23, Cohen d =0.33 (not significant, medium effect). Etomoxir: P =0.64, Cohen d = –0.16 (not significant, small effect). TGF-β1+etomoxir: P =0.76, Cohen d =0.24 (not significant, negligible effect). BSA: TGF-β1 vs control: P =0.006, Cohen d =−2.97 (significant, extremely large effect). Palm-BSA: TGF-β1 vs control: P <0.001, Cohen d =−10.95 (significant, extremely large effect). Etomoxir vs control: P =0.002, Cohen d =−3.93 (significant, extremely large effect). TGF-β1+etomoxir vs control: P =0.003, Cohen d =−3.03 (significant, extremely large effect). I , Rate of FAO after TGF-β1 treatment in HUVECs and HemECs (n=4 biological replicates). The paired 2-tailed Student t test was performed. HemECs: P =0.038, Cohen d =−1.75 (significant, large effect). HUVECs: P =0.89, Cohen d =−0.08 (not significant, negligible effect).

Article Snippet: HemEC isolation from proliferating IHs was performed as described previously., Primary human umbilical vein ECs (HUVECs) were obtained from the American Type Culture Collection (United States).

Techniques: Staining, Derivative Assay, Positive Control, Over Expression, Control, Western Blot, Expressing

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: TGF-β1 Promotes Angiogenesis via Endothelial-to-Mesenchymal Transition in Infantile Hemangioma

doi: 10.1161/ATVBAHA.125.322793

Figure Lengend Snippet: CPT1A KD (CPT1A [carnitine palmitoyltransferase 1A] knockdown) stimulates hemangioma-derived endothelial cell (HemEC) migration, invasion, and angiogenesis. A , Transwell migration assay results showing the migration ability of HemECs and human umbilical vein endothelial cells (HUVECs) after CPT1A KD , CPT1A KD +L-palmitoylcarnitine, and CPT1A OE +TGF-β1 OE (TGF-β1 [transforming growth factor β1] overexpression; n=4 biological replicates). Scale bar, 100 µm. The unpaired 2-tailed Student t tests were performed. P values were adjusted using the Benjamini-Hochberg correction. HUVECs: CPT1A KD vs control: P =0.10, Cohen d =1.95 (not significant, large effect). CPT1A KD +L-palmitoylcarnitine vs CPT1A KD : P =0.038, Cohen d =−2.75 (significant, extremely large effect). TGF-β1 OE +CPT1A OE vs CPT1A KD +L-palmitoylcarnitine: P =0.10, Cohen d =1.74 (not significant, large effect). HemECs: CPT1A KD vs control: P <0.001, Cohen d =18.11 (significant, extremely large effect). CPT1A KD +L-palmitoylcarnitine vs CPT1A KD : P <0.001, Cohen d =−14.42 (significant, extremely large effect). TGF-β1 OE +CPT1A OE vs CPT1A KD +L-palmitoylcarnitine: P =0.13, Cohen d =−1.53 (not significant, large effect). B , Transwell invasion assay results showing the invasion ability of HemECs and HUVECs after CPT1A KD , CPT1A KD +L-palmitoylcarnitine, or CPT1A OE +TGF-β1 OE (n=4 biological replicates). Scale bar, 100 µm. The unpaired 2-tailed Student t tests were performed. P values were adjusted using the Benjamini-Hochberg correction. HUVECs: CPT1A KD vs control: P =0.19, Cohen d =1.29 (not significant, extremely large effect). CPT1A KD +L-palmitoylcarnitine vs CPT1A KD : P =0.99, Cohen d =0 (not significant, no effect). TGF-β1 OE +CPT1A OE vs CPT1A KD +L-palmitoylcarnitine: P =0.83, Cohen d =0.19 (not significant, small effect). HemECs: CPT1A KD vs control: P =0.005, Cohen d =4.30 (significant, extremely large effect). CPT1A KD +L-palmitoylcarnitine vs CPT1A KD : P =0.005, Cohen d =−4.31 (significant, extremely large effect). TGF-β1 OE +CPT1A OE vs CPT1A KD +L-palmitoylcarnitine: P =0.16, Cohen d =1.33 (not significant, large effect). C , Tube formation assay showing the angiogenic ability of HemECs in vitro after CPT1A KD , CPT1A KD +L-palmitoylcarnitine, and CPT1A OE +TGF-β1 OE (n=4 biological replicates). Scale bar, 100 µm. D , Vessel formation results showing the angiogenic ability of HemECs in vivo after CPT1A KD , CPT1A KD +L-palmitoylcarnitine, and CPT1A OE +TGF-β1 OE (n=4 biological replicates). Scale bar, 50 µm. The unpaired 2-tailed Student t tests were performed. P values were adjusted using the Benjamini-Hochberg correction. Total length: HUVECs: CPT1A KD vs control: P =0.90, Cohen d =0.11 (not significant, small effect). CPT1A KD +L-palmitoylcarnitine vs CPT1A KD : P =0.18, Cohen d =1.31 (not significant, large effect). TGF-β1 OE +CPT1A OE vs CPT1A KD +L-palmitoylcarnitine: P =0.32, Cohen d =−0.93 (not significant, large effect). HemECs: CPT1A KD vs control: P =0.003, Cohen d =2.73 (significant, extremely large effect). CPT1A KD +L-palmitoylcarnitine vs CPT1A KD : P =0.013, Cohen d =−3.53 (significant, extremely large effect). TGF-β1 OE +CPT1A OE vs CPT1A KD +L-palmitoylcarnitine: P =0.84, Cohen d =0.17 (not significant, small effect). Number of branch sites. HUVECs: CPT1A KD vs control: P =0.91, Cohen d =0.10 (not significant, small effect). CPT1A KD +L-palmitoylcarnitine vs CPT1A KD : P =0.71, Cohen d= −0.33 (not significant, medium effect). TGF-β1 OE +CPT1A OE vs CPT1A KD +L-palmitoylcarnitine: P =0.85, Cohen d =0.16 (not significant, small effect). HemECs: CPT1A KD vs control: P =0.001, Cohen d =3.35 (significant, extremely large effect). CPT1A KD +L-palmitoylcarnitine vs CPT1A KD : P =0.013, Cohen d =−6.40 (significant, extremely large effect). TGF-β1 OE +CPT1A OE vs CPT1A KD +L-palmitoylcarnitine: P =0.66, Cohen d =−0.38 (not significant, small effect). D , Vessel formation results showing the angiogenic ability of HemECs in vivo after CPT1A KD , CPT1A KD +L-palmitoylcarnitine, and CPT1A OE +TGF-β1 OE (n=4 biological replicates). Scale bar, 50 µm. The unpaired 2-tailed Student t tests with the Welch correction were performed. CPT1A KD vs control: P <0.001, Cohen d =8.90 (significant, extremely large effect). CPT1A KD +L-palmitoylcarnitine vs CPT1A KD : P =0.014, Cohen d =−3.81 (significant, extremely large effect). TGF-β1 OE +CPT1A OE vs CPT1A KD +L-palmitoylcarnitine: P =0.72, Cohen d =0.25 (not significant, negligible to small effect).

Article Snippet: HemEC isolation from proliferating IHs was performed as described previously., Primary human umbilical vein ECs (HUVECs) were obtained from the American Type Culture Collection (United States).

Techniques: Knockdown, Derivative Assay, Migration, Transwell Migration Assay, Over Expression, Control, Transwell Invasion Assay, Tube Formation Assay, In Vitro, In Vivo

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: TGF-β1 Promotes Angiogenesis via Endothelial-to-Mesenchymal Transition in Infantile Hemangioma

doi: 10.1161/ATVBAHA.125.322793

Figure Lengend Snippet: CPT1A KD (CPT1A [carnitine palmitoyltransferase 1A] knockdown) suppresses hemangioma-derived endothelial cell (HemEC) autophagy. A , Transmission electron microscopy (TEM) results showing changes in autophagy after CPT1A KD , CPT1A KD +L-palmitoylcarnitine, and CPT1A OE +TGF-β1 OE (TGF-β1 [transforming growth factor β1] overexpression) in HemECs and human umbilical vein endothelial cells (HUVECs; n=4 biological replicates). Scale bar, 1 µm. B , Autophagic vesicles per field (n=4 biological replicates). The unpaired 2-tailed Student t tests were performed. P values were adjusted using the Benjamini-Hochberg correction. HUVECs: CPT1A KD vs control: P =0.33, Cohen d =0.83 (not significant, large effect). CPT1A KD +L-palmitoylcarnitine vs CPT1A KD : P =0.11, Cohen d =−1.54 (not significant, large effect). TGF-β1 OE +CPT1A OE vs CPT1A KD +L-palmitoylcarnitine: P =0.06, Cohen d =1.89 (not significant, large effect). HemECs: CPT1A KD vs control: P =0.004, Cohen d =−3.24 (significant, extremely large effect). CPT1A KD +L-palmitoylcarnitine vs CPT1A KD : P <0.001, Cohen d =9.71 (significant, extremely large effect). TGF-β1 OE +CPT1A OE vs CPT1A KD +L-palmitoylcarnitine: P =0.017, Cohen d =−2.55 (significant, large effect). C , Immunoblot analysis of TGF-β1, CPT1A, and AMPK (5’-monophosphate [AMP]–activated protein kinase; n=3 biological replicates).

Article Snippet: HemEC isolation from proliferating IHs was performed as described previously., Primary human umbilical vein ECs (HUVECs) were obtained from the American Type Culture Collection (United States).

Techniques: Knockdown, Derivative Assay, Transmission Assay, Electron Microscopy, Over Expression, Control, Western Blot

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: TGF-β1 Promotes Angiogenesis via Endothelial-to-Mesenchymal Transition in Infantile Hemangioma

doi: 10.1161/ATVBAHA.125.322793

Figure Lengend Snippet: Changes in the expression of endothelial and mesenchymal markers after R(+) propranolol or S(−) propranolol treatment in hemangioma-derived endothelial cells (HemECs) and human umbilical vein endothelial cells (HUVECs). A , Immunofluorescence images showing the expression of endothelial and mesenchymal markers before and after R(+) propranolol or S(−) propranolol treatment in HemECs and HUVECs. Scale bar, 100 µm (n=4 biological replicates). B , Immunofluorescence analysis showing the mean fluorescence intensity of endothelial and mesenchymal markers before and after R(+) propranolol or S(−) propranolol treatment in HemECs (n=4 biological replicates). The Dunnett multiple comparisons test was used. R(+) propranolol vs control: CD31: P <0.001, Cohen d =16.23 (significant, extremely large effect). VE-cadherin (vascular endothelial cadherin): P <0.001, Cohen d =11.58 (significant, extremely large effect). α-SMA (α-smooth muscle actin): P <0.001, Cohen d =−20.50 (significant, extremely large effect). COL1A1 (collagen type I alpha 1 chain): P =0.001, Cohen d =−11.23 (significant, extremely large effect). S(−) propranolol vs control: CD31: P =0.20, Cohen d =0.99 (not significant, large effect). VE-cadherin: P =0.07, Cohen d =2.62 (not significant, very large effect). α-SMA: P =0.75, Cohen d =0.21 (not significant, small effect). COL1A1: P =0.88, Cohen d =0.11 (not significant, negligible effect). C , Immunofluorescence analysis showing the mean fluorescence intensity of endothelial and mesenchymal markers before and after R(+) propranolol or S(−) propranolol treatment in HUVECs (n=4 biological replicates). The Dunnett multiple comparisons test was used. R(+) propranolol vs control: CD31: P =0.46, Cohen d =0.54 (not significant, medium effect). VE-cadherin: P =0.24, Cohen d =−0.94 (not significant, large effect). α-SMA: P =0.65, Cohen d =0.33 (not significant, small effect). COL1A1: P =0.52, Cohen d =0.48 (not significant, small effect). S(−) propranolol vs control: CD31: P =0.10, Cohen d =−1.36 (not significant, very large effect). VE-cadherin: P =0.79, Cohen d =1.90 (not significant, very large effect). α-SMA: P =0.93, Cohen d =−0.07 (not significant, negligible effect). COL1A1: P =0.87, Cohen d =0.1 (not significant, negligible effect).

Article Snippet: HemEC isolation from proliferating IHs was performed as described previously., Primary human umbilical vein ECs (HUVECs) were obtained from the American Type Culture Collection (United States).

Techniques: Expressing, Derivative Assay, Immunofluorescence, Fluorescence, Control