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Smooth muscle growth supplement

Manufactured by Thermo Fisher Scientific
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The Smooth Muscle Growth Supplement is a laboratory product designed to support the growth and development of smooth muscle cells in vitro. It provides essential nutrients and growth factors to facilitate the proliferation and differentiation of smooth muscle cell cultures. The core function of this product is to maintain and enhance the growth of smooth muscle cells in a controlled laboratory setting.

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Lab products found in correlation

Medium 231, by Thermo Fisher Scientific (12 mentions) Penicillin, by Thermo Fisher Scientific (7 mentions) Streptomycin, by Thermo Fisher Scientific (7 mentions) Hasmc, by Thermo Fisher Scientific (6 mentions) Low serum growth supplement, by Thermo Fisher Scientific (4 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (3 mentions) Losartan, by Bio-Techne (2 mentions) Pd123319, by Bio-Techne (2 mentions) Hcasmc, by Thermo Fisher Scientific (2 mentions) Smooth muscle differentiation supplement, by Thermo Fisher Scientific (2 mentions) Pcs 100 012tm, by Thermo Fisher Scientific (2 mentions) Trypsin edta, by Thermo Fisher Scientific (2 mentions) Smooth muscle cell growth medium, by Thermo Fisher Scientific (2 mentions) Penicillin streptomycin, by Merck Group (2 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Hpasmc, by Lonza (1 mentions) Sodium l ascorbate, by Merck Group (1 mentions) Dihydrotestosterone dht, by Merck Group (1 mentions) Flutamide, by Selleck Chemicals (1 mentions) Recombinant mouse tgf β, by R&D Systems (1 mentions)

Close spelling variants of this product

Growth supplement (20 citations) Smooth muscle cell growth supplement (5 citations) Smooth Muscle Growth Supplement (SMGS, (4 citations)

43 protocols using smooth muscle growth supplement

1

Cultivation of Human Coronary Artery Smooth Muscle Cells

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Human coronary artery smooth muscle cells (HCASMCs) from three donors were procured from GIBCO (Life Technologies, CA, USA) and LONZA (Walkersville MD, USA). The cells were pre-characterized for smooth muscle cell specific markers and for their compatible (<5% variation ) growth response to fetal calf serum (FCS; 2.5%). Cells were cultured in M231 culture medium containing smooth muscle growth supplement (Life Technologies, CA, USA) and under standard tissue culture conditions as described previously43 (link). HCASMCs in 3rd to 5th passage were used for the growth and molecular assays.
Dubey R.K., Fingerle J., Gillespie D.G., Mi Z., Rosselli M., Imthurn B, & Jackson E.K. (2015). Adenosine Attenuates Human Coronary Artery Smooth Muscle Cell Proliferation By Inhibiting Multiple Signaling Pathways that Converge on Cyclin D. Hypertension, 66(6), 1207-1219.
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2

Culturing Human Pulmonary Artery Smooth Muscle Cells

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HPASMCs were obtained commercially (Lonza, Walkersville, MD) and cultured in Medium 231 supplemented with Smooth Muscle Growth Supplement (Life technologies, Grand Island, NY) at 37 °C under 5% CO244 (link). In all experiments, HPASMCs were studied before passage 6 prior to any change in cell morphology.
Dong Y.N., Hsu F.C., Koziol-White C.J., Stepanova V., Jude J., Gritsiuta A., Rue R., Mott R., Coulter D.A., Panettieri RA J.r., Krymskaya V.P., Takano H., Goncharova E.A., Goncharov D.A., Cines D.B, & Lynch D.R. (2021). Functional NMDA receptors are expressed by human pulmonary artery smooth muscle cells. Scientific Reports, 11, 8205.
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3

Aortic Smooth Muscle Cell Culture

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Primary human aortic smooth muscle cells were purchased from the American Type Culture Collection (ATCC; PCS-100-012) and cultured in Medium 231 (Life Technologies), 500 ml of which was supplemented with 25 ml of growth supplement (smooth muscle growth supplement; Life Technologies) and sodium l-ascorbate (50 μg/ml; Sigma-Aldrich) for extracellular matrix secretion. The cells were seeded at a density of 1.25 × 104 per ml and used within the first three passages. The glass and PDMS surfaces were coated with collagen before seeding. The PDMS surfaces were oxygen plasma cleaned at 200 W for 10 min immediately before seeding collagen to make the surface hydrophilic.
Nair V., Yi J., Isheim D., Rotenberg M., Meng L., Shi F., Chen X., Gao X., Prominski A., Jiang Y., Yue J., Gallagher C.T., Seidman D.N, & Tian B. (2020). Laser writing of nitrogen-doped silicon carbide for biological modulation. Science Advances, 6(34), eaaz2743.
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4

Isolation and Characterization of PAH-Derived PASMC

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PAH PASMC were isolated from PAH patients. Normal PASMC were isolated from control subjects or purchased from Lonza or Cell Applications Inc. PASMC lines were studied within 6 passages. PASMC were grown in Medium 231 supplemented with Smooth Muscle Growth Supplement (SMGS, Life Technologies, Carlsbad, CA, USA). Experiments were performed on 6 normal PASMC lines (50% Female, Mean age: 50.7 years) and 8 PAH PASMC lines (28% Female, Mean age: 41.2 years). Blood Outgrowth Endothelial Cells (BOEC) were isolated and cultured from PAH patients and normal individuals (Demographics in Supplementary Table 1) as previously described20 (link). Briefly blood was collected from normal individuals (n=5) and PAH patients (n=6) by venipuncture. Peripheral blood mononuclear cell (PBMC) was isolated from the whole blood by density gradient centrifugation. PBMC were cultured in BOEC generation medium for 7–14 days for the appearance of outgrowth colonies.
Chen K.H., Dasgupta A., Lin J., Potus F., Bonnet S., Iremonger J., Fu J., Mewburn J., Wu D., Dunham-Snary K., Theilmann A.L., Jing Z.C., Hindmarch C., Ormiston M.L., Lawrie A, & Archer S.L. (2018). Epigenetic dysregulation of the Drp1 binding partners MiD49 and MiD51 increases mitotic mitochondrial fission and promotes pulmonary arterial hypertension: mechanistic and therapeutic implications. Circulation, 138(3), 287-304.
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5

Aortic SMCs Characterization Protocols

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Aortic SMCs were derived from the ASC of 16‐week‐old Fbn1C1039G/+ male and female mice, as previously described.18 Medium 231 and smooth muscle growth supplement were purchased from Life Technologies (Foster City, CA). SMC (2.0×105 cells/mL) were cultured to confluence in SMC media and starved with serum‐free medium for 24 hours before the treatment. SMC were treated with either dihydrotestosterone (DHT) (10 nmol/L) (Sigma‐Aldrich), flutamide (1 μmol/L) (Selleckchem, Houston, TX), mouse recombinant TGF‐β (5 ng/mL) (R&D Systems, Minneapolis, MN), or vehicle control for 24 hours for p‐ERK1/2 protein assay, 48 hours for MMP activity assay and 24 and 48 hours for androgen receptor protein assay. For p‐Smad2 protein assay, SMC were treated with mouse recombinant TGF‐β (5 ng/mL) or vehicle control for 1 hour after pretreatment with DHT, flutamide, or vehicle control for 24 hours. When applicable, flutamide was added into the media 3 hours before DHT treatment to block androgen receptor activation. Experiments included n=5 to 6 per group.
Tashima Y., He H., Cui J.Z., Pedroza A.J., Nakamura K., Yokoyama N., Iosef C., Burdon G., Koyano T., Yamaguchi A, & Fischbein M.P. (2020). Androgens Accentuate TGF‐β Dependent Erk/Smad Activation During Thoracic Aortic Aneurysm Formation in Marfan Syndrome Male Mice. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 9(20), e015773.
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6

Culturing and Treating Cardiac Cell Lines

Cited 1 time
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Mouse atrial cardiomyocyte HL-1 cells were a gift from Dr. William Claycomb at Louisiana State University Medical Center. HL-1 cells were cultured at 37°C and 5% CO2 on surfaces pre-treated with 12.5 μg/ml bovine fibronectin in 0.02% gelatin solution and grown in Claycomb medium supplemented with Fetal Bovine Serum (10%) (both from Sigma-Aldrich, St. Louis, MO), Penicillin/Streptomycin (100U/ml: 100mg/ml), Norepinephrine (100U/ml) and L-Glutamine (2mM) (GIBCO-Invitrogen) as described previously [38 (link), 41 (link)]. hCAVSMCs were purchased from GIBCO-Invitrogen Cell culture (Carlsbad, CA) and were cultured according to manufacturer’s instructions at 37°C and 5% CO2 in Medium 231 supplemented with Smooth Muscle Growth Supplement (SMGS, Life Technologies, Cat. No. S-007-25). β-blockers Aten, Met and Car and Ang II were purchased from Sigma-Aldrich, and Neb was a gift from Forest Laboratories Inc. (New York). Losartan (AT1R inhibitor), PD123319 (AT2R inhibitor), CGP42112a (partial AT2R agonist), SR59230A (β-3 AR specific blocker), U73122 (Phospholipase C inhibitor) and isoproterenol (β-AR activator) were purchased from Tocris Bioscience (Bristol, UK); NP-6A4 was a gift from Novopyxis Inc. (Boston).
Mahmood A, & Pulakat L. (2015). Differential Effects of β-Blockers, Angiotensin II Receptor Blockers, and a Novel AT2R Agonist NP-6A4 on Stress Response of Nutrient-Starved Cardiovascular Cells. PLoS ONE, 10(12), e0144824.
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7

Cultivation of Patient-Derived VSMC Cells

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Patient‐derived VSMC cell lines: Umb‐,29 Pla‐ and CerVSMC and human foreskin fibroblasts were cultured as described previously.35, 36 Umbilical artery smooth muscle cells (UASMC, Lonza) were cultured in smooth muscle cell medium BulletKit according to the manufacturer's instructions or cultured in M 231 medium with smooth muscle growth supplement (Life Technologies). Human aortic ECs (Life Technologies) were cultured in M 200 medium with low serum growth supplement (ThermoFisher Scientific).
Panahi M., Yousefi Mesri N., Samuelsson E., Coupland K.G., Forsell C., Graff C., Tikka S., Winblad B., Viitanen M., Karlström H., Sundström E, & Behbahani H. (2018). Differences in proliferation rate between CADASIL and control vascular smooth muscle cells are related to increased TGFβ expression. Journal of Cellular and Molecular Medicine, 22(6), 3016-3024.
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8

Exosome Secretion Quantification

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Medium RPMI 1640, Medium 231, Smooth Muscle Growth Supplement (SMGS), glutamine, antibiotics, HEPES, and phosphate buffered saline (PBS) were obtained from Life Technologies Corporation (Mulgrave, Victoria, Australia). CD63 ELISA kits were obtained from SBI (ExoELISA™, System Biosciences, Mountain View, CA).
Salomon C., Yee S., Scholz-Romero K., Kobayashi M., Vaswani K., Kvaskoff D., Illanes S.E., Mitchell M.D, & Rice G.E. (2014). Extravillous trophoblast cells-derived exosomes promote vascular smooth muscle cell migration. Frontiers in Pharmacology, 5, 175.
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9

Cultured Cell Line Protocols for Vascular Research

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All cells were grown at 37°C and 5% CO2. hCAVSMCs (GIBCO – Invitrogen Cell culture, Carlsbad, CA, United States) were cultured in Medium 231 supplemented with Smooth Muscle Growth Supplement (Life Technologies). hCAECs (Cell Applications, Inc.) were cultured in MesoEndo Cell Growth Medium (Cell Applications, Inc.). Human Umbilical Vein Endothelial Cells (hUVECs – GIBCO-Invitrogen Cell culture, Carlsbad, CA, United States) were cultured in Medium 200 supplemented with Low Serum Growth Supplement (Life Technologies). PD123319 (AT2R antagonist) and Losartan (AT1R antagonist) were purchased from Tocris Bioscience (Bristol, United Kingdom). NP-6A4 was a gift from Novopyxis Inc. (Boston, MA, United States). All experiments were performed on cells between passage number three and six. For collection, cells were rinsed twice with ice-cold PBS, scraped, centrifuged at 180 × g, and flash frozen with liquid nitrogen for subsequent assays.
Toedebusch R., Belenchia A, & Pulakat L. (2018). Cell-Specific Protective Signaling Induced by the Novel AT2R-Agonist NP-6A4 on Human Endothelial and Smooth Muscle Cells. Frontiers in Pharmacology, 9, 928.
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10

Cell Culture Protocols for Respiratory Research

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Calu-3 AEC (American Type Culture Collection, ATCC, (Manassas, VA, USA) were cultured as previously described [25 ]. Human non-neoplastic bronchial epithelial cells, BEAS-2B, were purchased from ATCC and cultured in bronchial epithelial cell growth medium (Lonza/Clonetics Corporation, Basel, Switzerland), following ATCC instructions. LA-4 adenoma cells were cultured in F–12K medium (Gibco, Waltham, MA, USA) supplemented with 15% FBS. A549 cells (ATCC) were maintained in Dulbecco’s Modified Eagle Medium (Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% FBS. PASMC were grown in Medium 231 supplemented with smooth muscle growth supplement (Life Technologies, Carlsbad, CA, USA). HPAEC (Lonza) were purchased and cultured in endothelial growth medium (EGM)-2 supplemented with the EGM-2 Bullet Kit (Lonza).
Archer S.L., Dasgupta A., Chen K.H., Wu D., Baid K., Mamatis J.E., Gonzalez V., Read A., Bentley R.E., Martin A.Y., Mewburn J.D., Dunham-Snary K.J., Evans G.A., Levy G., Jones O., Al-Qazazi R., Ring B., Alizadeh E., Hindmarch C.C., Rossi J., Lima P.D., Falzarano D., Banerjee A, & Colpitts C.C. (2022). SARS-CoV-2 mitochondriopathy in COVID-19 pneumonia exacerbates hypoxemia. Redox Biology, 58, 102508.
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