Human coronary artery endothelial cells (HCAEC) and human coronary artery smooth muscle cells (HCASMC) were purchased from Lonza. For HCASMC, donors included a 12 year old male, a 56 year old female, and a 30 year old male, while HCAEC donors were a 21 year old male and a 30 year old male. HCAEC were cultured in EBM-2 basal media supplemented with EGM-2MV SingleQuot factors (HCAEC complete media, Lonza). HCASMC were cultured in SMBM basal media supplemented with SmGM-2 SingleQuot factors (HCASMC complete media, Lonza). All cells were grown in a humidified incubator in 5% CO2 at 37 °C. Cells were used at passage five or six for all experiments. Serum reduced media for experiments to be performed in room air was prepared by diluting the appropriate complete media in low glucose DMEM (Hyclone, Fisher) at a ratio of 2:3 yielding a 2% serum media, and supplementing with 15 mmol/L HEPES.
Hcasmc
Manufactured by Lonza
Sourced in United States
HCASMC is a primary human cell line derived from the smooth muscle cells of the coronary artery. This cell line is designed for in vitro research purposes.
Automatically generated - may contain errors
Lab products found in correlation
Hcaec, by Lonza (9 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Penicillin streptomycin solution, by Thermo Fisher Scientific (2 mentions)
Smgm 2 singlequots, by Lonza (2 mentions)
Human umbilical vein endothelial cells (huvec), by Lonza (2 mentions)
Cc 3182, by Lonza (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Ebm 2, by Lonza (2 mentions)
Huvecs, by Lonza (2 mentions)
Basal medium, by Thermo Fisher Scientific (1 mentions)
Smooth muscle differentiation supplement, by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle medium, by Thermo Fisher Scientific (1 mentions)
Ebm 2 basal media, by Lonza (1 mentions)
Smbm basal medium, by Lonza (1 mentions)
Normal human dermal fibroblasts (nhdf), by Lonza (1 mentions)
Human recombinant tnfα, by R&D Systems (1 mentions)
Mouse recombinant tnfα, by R&D Systems (1 mentions)
Ifn γ, by Thermo Fisher Scientific (1 mentions)
Il 1β, by R&D Systems (1 mentions)
Ifn γ, by R&D Systems (1 mentions)
22 protocols using hcasmc
1
Culturing Coronary Artery Cells from Diverse Donors
2
Stimulation of Coronary Cells
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Human coronary artery endothelial cells (HCAECs) and human coronary artery smooth muscle cells (HCASMCs) were purchased from Lonza. HCAECs and HCASMCs were grown in Endothelial Cell Basal Medium‐2 (EBM‐2; Lonza) supplemented with EGM‐2 MV SingleQuots (Lonza) and in Smooth Muscle Cell Basal Medium (Lonza) supplemented with SmGM‐2 SingleQuots (Lonza), respectively, according to the manufacturer's instructions. After serum starvation for 24 hours, cells were stimulated with 50 ng/mL of vascular endothelial growth factor (VEGF) (R&D Systems) for 0.5 or 24 hours, 0.5 μmol/L insulin for 0.5 hours, 100 μmol/L H2O2 for 1 hour followed by 23‐hour incubation in normal culture medium after washing, 0 to 200 nmol/L recombinant FABP4, or 10 μg/mL anti‐FABP4 antibody in the medium supplemented with 0.5% BSA. The doses of reagents and incubation periods varied according to the experimental protocol. Each experiment was performed in at least triplicate.
3
Culturing Human Cell Lines for Notch1 Signaling
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Human embryonic kidney
(HEK) 293T cells stably expressing full-length Notch1 (HEK293 FLN1)
were cultured under puromycin selection (1 μg/mL) in complete
medium consisting of Dulbecco’s modified Eagle medium (Gibco)
supplemented with 5 vol % heat inactivated fetal bovine serum (Invitrogen)
and 1 vol % penicillin–streptomycin solution (Invitrogen),
at 37 °C and 5% CO2 in a humidified atmosphere. HEK293
FLN1 were seeded at a concentration of 1.0 × 105 cells/cm2.
Primary human coronary artery smooth muscle cells
(HCASMC, Lonza) were purchased at passage 3 and experiments were carried
out with cells at passage 5 or 6. Cells were cultured in 231 basal
medium (Gibco) supplemented with 5 vol % smooth muscle growth supplement
(SMGS, Gibco) and 1 vol % penicillin–streptomycin solution
(Invitrogen), at 37 °C and 5% CO2 in a humidified
incubator. As a control for the expression of α-smooth muscle
actin in the differentiated state, the basal medium was supplemented
with 1 vol % smooth muscle differentiation supplement (Gibco) instead
of SMGS. HCASMC was passaged at 80% confluency and seeded at a concentration
of 5 × 103 cells/cm2.
(HEK) 293T cells stably expressing full-length Notch1 (HEK293 FLN1)
were cultured under puromycin selection (1 μg/mL) in complete
medium consisting of Dulbecco’s modified Eagle medium (Gibco)
supplemented with 5 vol % heat inactivated fetal bovine serum (Invitrogen)
and 1 vol % penicillin–streptomycin solution (Invitrogen),
at 37 °C and 5% CO2 in a humidified atmosphere. HEK293
FLN1 were seeded at a concentration of 1.0 × 105 cells/cm2.
Primary human coronary artery smooth muscle cells
(HCASMC, Lonza) were purchased at passage 3 and experiments were carried
out with cells at passage 5 or 6. Cells were cultured in 231 basal
medium (Gibco) supplemented with 5 vol % smooth muscle growth supplement
(SMGS, Gibco) and 1 vol % penicillin–streptomycin solution
(Invitrogen), at 37 °C and 5% CO2 in a humidified
incubator. As a control for the expression of α-smooth muscle
actin in the differentiated state, the basal medium was supplemented
with 1 vol % smooth muscle differentiation supplement (Gibco) instead
of SMGS. HCASMC was passaged at 80% confluency and seeded at a concentration
of 5 × 103 cells/cm2.
4
Culturing Human Coronary Artery Smooth Muscle Cells
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Primary human coronary artery smooth muscle cells (HCASMC) were purchased from Lonza and cultured in growth media containing SmBM Basal Medium (CC-3181, Lonza) and SmGM-2 SingleQuots supplements (CC-4149) required for growth of smooth muscle cells. THP1 monocytic cell line (DSMZ, Cat.No: ACC16) was cultured in RPMI 1640 medium supplemented with 10% (v/v) FCS, 100 U/ml penicillin and 100 mg/ml streptomycin.
5
Culturing Primary Human Vascular Cells
6
Cytokine-Stimulated Vascular Smooth Muscle Cell Model
7
Coronary Artery Cell Culture Protocols
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Human coronary artery endothelial cells (HCAEC) and human coronary artery smooth muscle cells (HCASMC) were obtained from Lonza Inc, Allendale, NJ. HACEC were cultured in optimized EGM‐2 medium (cat# CC‐3162, Clonetics® , Lonza) and HCASMC were grown in optimized SmGM‐2 media (cat# CC‐3182, Clonetics® , Lonza). HCAEC passages 5–14 and HCASMC passages 4–10 were used in experiments and culture medium was renewed every 2 days. Cells were grown on fibronectin‐coated 10‐cm dishes covered in parafilm and maintained in a standard humidified incubator at 37°C in a 5% CO2 atmosphere. Cells were treated with/without experimental agents as indicated (Ethanol 5–50 mM; DLL4, 3 μg/ml; DAPT, 20 μM). Under these conditions, there was little change (<10%) in ethanol concentration (determined by diagnostic kit from Sigma‐Aldrich) after 24‐h incubation +/− HCAEC or HCASMC indicating no significant metabolism of ethanol by these cells.
8
Culturing Human Coronary Artery Smooth Muscle Cells
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Primary human coronary artery smooth muscle cells (HCASMC) were purchased from three different manufacturers, Lonza, PromoCell and Cell Applications and were cultured in complete smooth muscle basal media (Lonza, #CC-3182) according to the manufacturer's instructions. All experiments were performed with HCASMC between passages 5–8. HEK293 cells were maintained in DMEM containing high glucose, sodium pyruvate and L-glutamine supplemented with 10% FBS.
9
Culturing Primary Human Coronary Artery Smooth Muscle Cells
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Primary human coronary artery smooth muscle cells (hCASMC) were purchased from Lonza (#CC-2583) and cultured in M199 supplemented with 10% FBS, rhEGF (2.7 ng/ml), rhFGF (2 ng/ml), penicillin G (100 units/ml) and streptomycin sulfate (100 μg/ml). The cells were starved with starvation media containing 0.1% FBS and 0.1% bovine serum albumin (BSA) for 24 h prior to experimental treatments. Subconfluent monolayers of fourth to seventh passage hCASMCs were used in all experiment.
10
Isolation and Culture of Human Coronary Artery Smooth Muscle Cells
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Primary human coronary artery smooth muscle cells (HCASMC) were purchased from three different manufacturers, Lonza, PromoCell and Cell Applications and were cultured in complete smooth muscle basal media (Lonza, #CC-3182) according to the manufacturer's instructions. All experiments were performed with HCASMC between passages 4–7. Genotypes of HCASMC were determined as described above, and lots heterozygous at rs12190287 were used for all experiments. The A7r5 rat aortic SMC line was purchased from ATCC and cells were maintained in Dulbecco's modified Eagle medium (DMEM, Life Technologies, #11885-084) containing low glucose, sodium pyruvate and L-glutamine and supplemented with 10% fetal bovine serum (FBS). HeLa cells were maintained in DMEM containing high glucose, sodium pyruvate and L-glutamine supplemented with 10% FBS.
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