Human umbilical vein endothelial cells (HUVECs) and human aortic smooth muscle cells (HASMCs) were purchased from ScienCell Research Laboratories (Cat#8000, #6110, ScienCell), HUVECs were cultured in endothelial cells medium (Cat#1001, ScienCell) with 5% fetal bovine serum (FBS), 1% endothelial cells growth supplement (ECGS), and 1% penicillin/streptomycin; HASMCs were cultured in smooth muscle cell medium (SMCM, Cat. #1101, ScienCell, USA) with 2% FBS (Cat#1600-0044, Gibco), 1% SMCGS, and 1% penicillin and streptomycin. Experiments were performed using cells from passages 2–8. Human myeloid leukemia mononuclear cells (THP-1) obtained from Fudan University Institutes of Biomedical Sciences Cell Center (Shanghai, China) were cultured in Dulbecco's Modified Eagle Medium (DMEM, Cat#ZQ-100, Shanghai Zhong Qiao Xin Zhou Biotechnology) with 10% FBS and 1% penicillin/streptomycin. Isolation of RASMCs from the thoracic aortas of male Sprague–Dawley rats were performed as previously described (18 (link)). Cells were cultured in DMEM with 20% FBS and 1% penicillin/streptomycin. All types of cells were incubated in 5% CO2 at 37°C.
Hasmc
Manufactured by ScienCell
Sourced in United States
HASMCs are primary human aortic smooth muscle cells. They are isolated from the tunica media of the human aorta and are cryopreserved at the second passage to retain their original characteristics.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by ScienCell (8 mentions)
Smooth muscle cell medium, by ScienCell (7 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions)
Smcgs, by ScienCell (4 mentions)
Hasmc, by Thermo Fisher Scientific (4 mentions)
Trypsin edta solution, by Thermo Fisher Scientific (3 mentions)
Bright line, by Hausser Scientific (3 mentions)
Ang 2, by Merck Group (3 mentions)
Huvecs, by ScienCell (3 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions)
Trypsin edta, by Thermo Fisher Scientific (3 mentions)
Pdgf bb, by Merck Group (2 mentions)
Pdgf bb, by R&D Systems (2 mentions)
Hasmcs, by MedChemExpress (2 mentions)
Endothelial cell medium (ecm), by ScienCell (2 mentions)
Primary hasmcs, by ScienCell (2 mentions)
Smooth muscle cell growth supplement, by ScienCell (2 mentions)
5000 tension apparatus, by Flexcell (2 mentions)
Hcaec, by ScienCell (2 mentions)
Rnaimax, by Thermo Fisher Scientific (2 mentions)
52 protocols using hasmc
1
Isolation and Culture of Vascular Cells
2
Adhesion of Vascular Cell Types on SilkGraft
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Adult Human Coronary Artery Endothelial Cells (HCAECs), Human Aortic Smooth Muscle Cells (HASMCs), and Human Aortic Adventitial Fibroblasts (HAAFs) were provided by ScienCell Research Laboratories (Carlsbad, CA, USA). The supplier company guaranteed the cells characteristics we required via Cell Applications Inc. (San Diego, CA, USA).
For the adhesion studies, 2 × 104 human intravitally pre-stained cells were separately seeded onto SilkGraft samples. HCAECs were seeded onto the inner ES layer, whereas HASMCs or HAAFs were seeded onto the outer ES layer. In parallel, the three cell types were separately seeded on 2D polystyrene plates as controls. All the tests were performed in triplicate and repeated in three separate experiments. Cell cultures were kept in an incubator at 37°C in 95 vol% air plus 5 vol% CO2. The growth media used were: HCAECs, ready to use Endothelial Cell Medium; HAAFs, Fibroblast Medium; HASMCs, Smooth Muscle Cell Medium (all from ScienCell Research Laboratories, USA). Every 3 days the growth media were changed with fresh ones and the cell-conditioned media collected and stored at −80°C to be subsequently analyzed. The cultures were kept going for at least 20 consecutive days.
For the adhesion studies, 2 × 104 human intravitally pre-stained cells were separately seeded onto SilkGraft samples. HCAECs were seeded onto the inner ES layer, whereas HASMCs or HAAFs were seeded onto the outer ES layer. In parallel, the three cell types were separately seeded on 2D polystyrene plates as controls. All the tests were performed in triplicate and repeated in three separate experiments. Cell cultures were kept in an incubator at 37°C in 95 vol% air plus 5 vol% CO2. The growth media used were: HCAECs, ready to use Endothelial Cell Medium; HAAFs, Fibroblast Medium; HASMCs, Smooth Muscle Cell Medium (all from ScienCell Research Laboratories, USA). Every 3 days the growth media were changed with fresh ones and the cell-conditioned media collected and stored at −80°C to be subsequently analyzed. The cultures were kept going for at least 20 consecutive days.
3
Adenoviral Infection of Vascular Cells
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HASMCs were obtained from ScienCell Research Laboratories (Catalog #6,110), and HCASMCs and HCAECs were from PromoCell Research Laboratories (Catalog #C-12511, #C-12221); all the cells were P0 or P1 primary cells, not immortal cell lines. HCASMCs and HASMCs were cultured in SMC medium (ScienCell, 1101) while HCAECs were cultured in endothelial cell medium (ScienCell, 1001); they were used at passages 4–7. HASMC, HCASMC and HCAEC synchronization were achieved by starving the cells in SMC medium or endothelial cell medium for 24 h followed by Ad-GFP, Ad-SRSF1, Ad-Δ133p53, Ad-p53, Ad-Δ40p53 or Ad-KLF5 (Krüppel-like factor 5) at a multiplicity of infection of 50 or 100 (50 or 100 plaque-forming units per cell) for 48 h. The infection efficiency was assessed by western blot for each adenoviral vector.
4
Modulation of Vascular Smooth Muscle Cells by PTH
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Primary human aortic smooth muscle cells (HASMCs) were purchased from ScienCell Research Laboratories (Carlsbad, USA) and cultured as described previously [16 (link)]. Briefly, HASMCs were cultured at 37 °C in a humidified atmosphere with 5% CO2 in smooth muscle cell growth culture medium (1% smooth muscle cell growth supplement in basal medium with 2% FBS and 1% penicillin-streptomycin (ScienCell)). The medium was replaced every other day. All experiments used third to fifth passage cells. Human PTH fragment 1–34 was purchased from Sigma-Aldrich (P3796). HASMCs were treated with various concentrations (1 × 10−6, 1 × 10−7, or 1 × 10−8 mol/L) of PTH for different time (0, 3, 7, 10, or 14 days), HASMCs cultured in normal medium for the same days as control. Smooth muscle cell growth culture medium was used for all experiments.
5
Smooth Muscle Cell Culture and IS Treatment
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HASMCs (Lot. 6001) were obtained from ScienCell Research Laboratory, Carlsbad, CA. HASMCs were cultured in 5 % CO2 at 37°C incubator with smooth muscle cell medium containing 0.5 mmol/L phosphate and 1.6 mmol/L calcium (Catalog No. 1101; ScienCell Research Laboratory, Carlsbad, CA). DMEM (Catalog No.1013, Invitrogen, Carlsbad, CA) with 0.5% fetal bovine serum was used as starvation medium for short-term experiments to assess the role of IS. HASMCs cultures at ~70% confluence were synchronized under serum-free conditions for 48 h and then treated with IS at concentrations of 0, 200, 500, and 1000 μmol/L for 6 d. For inhibition experiments, 5-aza-2'-deoxycytidine (5Aza-2dc) at concentrations of 0, 0.1, 1, and 10 μmol/L were added to HASMCs treated with IS at a concentration of 1000 μmol/L for 6d. Each treatment was done in triplicates.
6
Culturing HUVEC and HASMC for TEBV
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Human umbilical vein endothelial cells (HUVECs) and human aortic smooth muscle cells (HASMCs) were obtained from the ScienCell Research Laboratories and cultured according to the established cell culturing protocols (Chen et al., 2018 (link)). HUVECs were cultured in endothelial cell medium (ScienCell), while HASMCs cultured in smooth muscle cell medium (ScienCell). When reaching 80% confluency, the cells were trypsinized (0.05% trypsin/EDTA; Thermo Fisher) and passaged. All cells were maintained in standard culture conditions (37 °C in humidified air with 5% CO2), and all cells were used within six to eight passages after reception to fabricate the TEBV.
7
Cell Culture and siRNA Transfection
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A7r5 was purchased from Cell Bank of Type Culture Collection of Chinese Academy of Sciences (Shanghai, China). HASMC was purchased from ScienCell Research Laboratories (Carlsbad, CA, United States). These cell lines were cultured in Dulbecco’s modified Eagle’s medium (4.5 g/L glucose, DMEM) containing 10%fetal bovine serum (FBS) as well as 100 μg/mL penicillin/streptomycin at 37°C in a humidified atmosphere containing 5% CO2. Cells were seeded in 60 mm dishes at an initial density of 1 × 105 cells/well and grown to approximately 80% confluence. For the siRNA transfection experiments, VSMC were grown to 60% confluence and AMPK α1/2 siRNA mix or negative control (NC) siRNA (Santa Cruz, CA, United States) were transfected using Lipofectamine 3000 (Invitrogen, CA, United States) according to the manufacturer’s instructions.
8
Modulating HASMC via ARF6 and MMP14
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Human Aortic Smooth Muscle Cells (HASMC) were obtained from ScienCell Research Laboratories (Carlsbad, CA), cultured in smooth muscle cell medium (ScienCell) according to the manufacturer’s instructions and maintained in a humidified incubator with 5% CO2 at 37 °C. All experiments were performed between passages 4–8. HASMC were infected in the presence of polybrene (8 μg/ml) with untargeted (control), ARF6 shRNA or MMP14 shRNA lentiviruses. Stable clones were selected using puromycin (5 μg/ml) and cells were used for experiments 72 h post-infection. HASMC were transfected with ARF6T27N pcDNA 3.0 or empty plasmid (Ctl) in the presence of Lipofectamine 3000 according to the manufacturer’s instructions. Cells were used for experiments 72 post-transfection.
9
Culturing Human Aorta Cells
10
Culture of Aortic Smooth Muscle and Macrophage Cells
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Human aortic Smooth muscle cell line (HA-SMC, ScienCell, 6110, San Diego, CA) was cultured in dedicated SMC conditional medium (ScienCell, 1101) supplemented with 2% of heat-inactivated fetal bovine serum (FBS, ScienCell, 0010), growth supplement (ScienCell, 1152), penicillin (100 U/ml) and streptomycin (100 μg/ml) (ScienCell, 0503) at 37 °C in a humidified 5% CO2 incubator. All cells used in this study were between passages 4 and 6. Human mononuclear macrophage cell line (THP-1, Chinese Academy of Science, TCHu 57, Shanghai, China) was cultured in 1640 medium (Gibco, RPMI 1640, 11875-093) supplemented with 10% FBS, penicillin (100 U/ml) and streptomycin (100 μg/ml, Gibco, 10378-016) at the same condition of culture.
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