Human umbilical vein endothelial cells (HUVEC), human aortic smooth muscle cells (HASMC) and human normal dermal fibroblasts (HNDFB) were purchased from Lonza, Inc. (Walkersville, MD, USA). HUVEC, HASMC and HNDFB were cultured in an appropriate medium, i.e., endothelial cell medium (ECM), smooth muscle cell medium (SMCM), fibroblast medium (FM), respectively, with growth supplement (Lonza). The cells were passaged every 3 days and were used within the third to fifth passage in this study. The cells were cultured on 0.5% gelatin-coated dishes (Techno Plastic Products, Sigma-Aldrich) and were maintained at 37°C in a humidified atmosphere containing 5% CO2. HUVEC were incubated with CellTracker Red (10μM) (Molecular Probes, Invitrogen Corp, Carlsbad, CA) for 30 minutes to allow visualization of the HUVEC in MCSs. Mixed cell suspensions (a total cell count of 2.5 x 104) composed of HUVEC (40%), HASMC (10%), and HNDFB (50%) was plated into each well of ultra-low attachment round-shaped 96-U-well plates (Sumitomo Bakelite, Tokyo, Japan) filled with triple mixed media of ECM, SMCM, and FM with a ratio of 1:1:1. After 24 hours, the cells aggregated to form a round shaped MCS. According to the automated measurement function equipped on our Bio-3D printer, the size of the MCS was 615.0±51.3 μm (mean ±SD) for 500 MCSs.
Hasmc
Manufactured by Lonza
Sourced in United States, Switzerland
HASMCs are primary human aortic smooth muscle cells used for in vitro research. They are non-transformed, non-immortalized cells isolated from the aorta of healthy human donors.
Automatically generated - may contain errors
Lab products found in correlation
Smgm 2, by Lonza (5 mentions)
Huvecs, by Lonza (5 mentions)
Biorevo bz 9000 microscope, by Keyence (4 mentions)
Ang 2, by Merck Group (3 mentions)
Thp1 monocytes, by Health Science Research Resources Bank (3 mentions)
Smgm 2 medium, by Lonza (2 mentions)
Fetal bovine serum (fbs), by Lonza (2 mentions)
Hasmc, by Merck Group (2 mentions)
Egm 2, by Lonza (2 mentions)
Huvecs, by American Type Culture Collection (2 mentions)
Hasmc, by American Type Culture Collection (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Tritc conjugated phalloidin, by Merck Group (1 mentions)
C1si spectral confocal microscope, by Nikon (1 mentions)
Hoechst 33258, by Thermo Fisher Scientific (1 mentions)
Celltracker red, by Thermo Fisher Scientific (1 mentions)
Growth supplement, by Lonza (1 mentions)
Hndfb, by Lonza (1 mentions)
Human umbilical vein endothelial cells (huvec), by Thermo Fisher Scientific (1 mentions)
30 protocols using hasmc
1
Multicellular Spheroid Formation Methodology
2
Asthmatic Airway Smooth Muscle Cell Culture Protocol
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Human ASM cells (HASMC) were purchased from Lonza, Walkersville, MD. Donors were three adult asthmatic subjects (Catalog number 194850, Lonza Inc., Switzerland) and three disease-free and non-smokers (Catalog number CC-2576, Lonza Inc., Switzerland). HASMC were cultured in vitro as previously described [20] (link), [21] (link). Briefly, HASMC were cultured in growth medium supplemented with 10% FBS (Bio Whittaker), and maintained in an incubator containing 5% CO2 in air at 37°C. After attaining ∼95% confluence, the cells were starved in unsupplemented Ham's F12 media for 24 hr. For all experiments cells were used at passage 2–4. HASMC were stimulated with either TSLP (10 ng/ml R&D Systems, Minneapolis, MN, USA) or dsRNA analogue (Poly I:C 10 ug/ml, polyinosine-polycytidylic acid; InvivoGen San Diego, CA, USA). Concentrations of these reagents were based on prior studies [21] (link), [22] . After exposure to drugs and control media, confluent HASM cells were stimulated for 0, 24 or 48 h by adding dsRNA or TSLP to the wells. Cell supernatant was collected at the different time points and maintained at −80°C until further analysis.
3
Differentiation of Human Aortic Smooth Muscle Cells
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Human aortic smooth muscle cells (HASMCs) were purchased from Lonza (Walkersville, MD, United States). HASMCs were grown in HASMC growth media [SmGM™-2 (Lonza) containing supplements and growth factors, including FGF-2 and EGF, in the kit (Lonza)]. HASMC cultures between passages two and five were used in all experiments. HASMCs were incubated with SmGM™-2 supplemented with 1% FBS for 7 days for differentiation.
4
Culturing Human Aortic Smooth Muscle Cells
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HASMC were obtained from Lonza (cc-2571). Cells were maintained in SmGM-2 medium (Lonza; cc-3181) supplemented with 20% FBS (Sigma) at 37°C in a humidified atmosphere with 5% CO2. HASMC of passage 5– 8 were used in the experiments.
5
Optimizing UII-Induced HASMC Proliferation
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Human aortic SMCs (HASMC) (Lonza, Walkersville, MD, USA) were cultured in growth media SmGM-2 (Lonza) in 5% fetal bovine serum at 37°C in a humidified 5% CO2 incubator. From the preliminary test for optimum concentration of UII, we found out 50 nM UII as optimal conc. for HASMC proliferation. After serum starvation for 24 h, cells were pretreated with UT antagonist or inhibitors for 30 min before UII treatment.
6
Nitinol Substrate Cellular Morphology
7
Vascular Smooth Muscle Cell Culture
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HASMC (Lonza Group, Basel, Switzerland) were cultured in Dulbecco's Modified Eagle Medium (Sigma-Aldrich, St. Louis, MO) containing 10% (v/v) fetal calf serum, 1% (v/v) L-glutamine and penicillin/streptomycin (100 U ml−1/100 μg ml−1) in a 5% CO2/95% humidified air incubator at 37 °C as described before.20 (link),21 (link) Cells were passaged with trypsin and all experiments were performed between passages 6 and 10. Cells were treated with either recombinant human klotho (1 nM) (R&D systems, Abingdon, UK) or AngII (100 or 200 nΜ) (Sigma-Aldrich, St. Louis, MO), for the indicated periods of time.
8
Culturing Human Aortic SMCs
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Human aortic SMCs (HASMC) were purchased from Lonza. Cells were cultured using Smooth Muscle Cell Growth Medium (SmGM) containing 5% fetal bovine serum with growth factors and antibiotics (Lonza, Cat no. cc-4149) in humidified incubator containing 5% CO2 at 37 °C. All experiments were performed with HASMCs at passages 5–7.
9
Rat and human aortic smooth muscle cell culture
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Rat and mouse aortic smooth muscle cells were cultured in growth medium (DMEM:F12, Gibco, 11320-033) supplemented with fetal bovine serum (10%, Corning, 35-015-CV), L-glutamine (1.6 mM, Gibco, 25030081), and penicillin-streptomycin (100 U/mL, Gibco, 15140122) in a humidified sterile incubator at 37°C with 5% CO2. Rat aortic smooth muscle cells constitutively expressing Myocd-LSD1 were generated by retroviral transduction as previously described (Liu et al., 2021 (link)). Prior to all stimulations and treatments, SMC were starved in a serum-free, insulin-free medium supplemented with L-glutamine (1.6 mM, Gibco, 25030081), L-ascorbic acid (0.2 mM, Sigma Aldrich, A4403), Apo-Transferrin (5 mg/ml, Sigma Aldrich, T5391) and Na Selenite (6.25 ng/ml, Sigma-Aldrich, S5261) for 24 to 48 h at 37°C with 5% CO2. Independent experiments were performed using SMC from constitutive passages. Human recombinant PDGF-BB (Sigma Aldrich, SRP3138) was reconstituted in 10 mM Acetic Acid at a final concentration of 30 ng/ml for treatment. SMC were treated with PDGF-BB for 24h before being harvested for analysis.
Human aortic smooth muscle cells (hASMCs, Lonza) were cultured in SmBMTM Basal Medium (CC-3181, Lonza) supplemented with SmGMTM-2 SingleQuotsTM (CC-4149, Lonza), 10% FBS and 100 U/ml each penicillin-streptomycin. Cells were starved in serum-free medium for 16 h before treatment.
Human aortic smooth muscle cells (hASMCs, Lonza) were cultured in SmBMTM Basal Medium (CC-3181, Lonza) supplemented with SmGMTM-2 SingleQuotsTM (CC-4149, Lonza), 10% FBS and 100 U/ml each penicillin-streptomycin. Cells were starved in serum-free medium for 16 h before treatment.
10
HASMC Migration Assay with AngII and KP-10
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HASMCs (Lonza) at passage 6 to 8 were seeded into 8‐well culture slides (3×103 cells/250 μL/well). Cells were incubated at 37°C in 5% CO2 for 3 to 5 hours in SmGM‐2 (Lonza) containing 5% FBS. HASMCs were then incubated for 15 hours with the indicated concentrations of angiotensin II (AngII) and/or KP‐10 in serum‐free SmGM. Cells were photographed at 10‐minute intervals. The average migration distance of 10 cells randomly selected in each dish was measured using a BIOREVO BZ‐9000 microscope (Keyence Corp, Osaka, Japan).19 , 20 , 21 , 22 , 23
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