Smgm 2 bulletkit

Manufactured by Lonza

Sourced in Switzerland, United States

The SmGM-2 BulletKit is a specialized laboratory equipment designed for the smooth and efficient cultivation of smooth muscle cells. It provides a comprehensive set of components required for the growth and maintenance of these cell types, including a growth medium, supplements, and other necessary reagents.

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Lab products found in correlation

Egm 2 bulletkit, by Lonza (7 mentions) Aosmc, by Lonza (4 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) Streptomycin, by Thermo Fisher Scientific (3 mentions) Penicillin, by Thermo Fisher Scientific (3 mentions) Bj 5ta, by American Type Culture Collection (2 mentions) 2mml glutamine, by Thermo Fisher Scientific (2 mentions) Nhost, by Lonza (2 mentions) Nucleofector system, by Lonza (2 mentions) Gfp trap gfp binding protein, by Thermo Fisher Scientific (2 mentions) Ndhfkit, by Lonza (2 mentions) Dulbecco modified eagle medium (dmem), by Euroclone (1 mentions) Penicillin, by Euroclone (1 mentions) Streptomycin, by Euroclone (1 mentions) Ebm basal medium, by Lonza (1 mentions) Egm singlequots kit, by Lonza (1 mentions) Human umbilical vein endothelial cells (huvec), by Lonza (1 mentions) Multiskan ascent plate reader, by Thermo Fisher Scientific (1 mentions) Premix wst 1 cell proliferation assay system, by Takara Bio (1 mentions) Huvecs, by Lonza (1 mentions)

Spelling variants of this product

SmGM-2 (60 citations) SmGM-2 BulletKit medium (11 citations) BulletKits (4 citations) SmGM-2 BulletKit media (4 citations) SmGM™-2 Smooth Muscle Cell Growth Medium -2 BulletKit™ (2 citations)

19 protocols using smgm 2 bulletkit

Culturing Colorectal Adenocarcinoma and Primary Intestinal Epithelial Cells

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Human colorectal adenocarcinoma Caco-2 cell line (ATCC HTB-37) was cultured in DMEM (Euroclone) supplemented with 10% fetal bovine serum (FBS, Gibco), 2 mM glutamine, 100 U/ml penicillin, 100 mg/ml streptomycin (Euroclone). Cell viability was estimated by Trypan blue (0.02%) exclusion (Sigma-Aldrich).
Primary human intestinal epithelial cells (Clonetics InEpC’s, Lonza) were grown in SmBM smooth muscle cell basal medium added with the supplements and growth factors insulin, hFGF-B, hEGF, FBS and gentamicin/amphotericin-B (SmGM-2 BulletKit, Lonza) at 33°C.

Aleandri M., Conte M.P., Simonetti G., Panella S., Celestino I., Checconi P., Marazzato M., Longhi C., Goldoni P., Nicoletti M., Barnich N., Palamara A.T., Schippa S, & Nencioni L. (2015). Influenza A Virus Infection of Intestinal Epithelial Cells Enhances the Adhesion Ability of Crohn’s Disease Associated Escherichia coli Strains. PLoS ONE, 10(2), e0117005.

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Culturing Human Cell Lines for Research

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Primary human foreskin fibroblasts (HFFs), immortalized human
fibroblasts (BJ5Ta and BR5, ATCC), human adenocarcinoma line MDA-MB-231, primary
human osteoblasts (NhOst, Lonza), and HEK 293FT cells were cultured in phenol
red-free DMEM (Hyclone) supplemented with 10% fetal bovine serum (Hyclone), 100
U/ml penicillin, 100 μg/ml streptomycin (Invitrogen), and 2 mM
L-glutamine (Invitrogen) at 37°C in 10% CO₂ in a humidified
incubator. Human umbilical vein endothelial cells (HUVECs) and human aortic
smooth muscle cells (AOSMCs, Lonza) were cultured in phenol red-free DMEM
(Hyclone) supplemented with 5% fetal bovine serum (Hyclone), insulin, hFGF, and
hEGF (Lonza, SMGM-2 BulletKit) at 37°C in 5% CO₂. The
following reagents were used in this study: rhodamine- and Alexa488-phallodin
(Invitrogen), cell-permeable C3 transferase (Cytoskeleton), blebbistatin and
okadaic acid (EMD), and GFP-TRAP GFP-binding protein (Chromotek). GFP-RhoQ63L
was transfected into cells with the Nucleofector system (Lonza) using the NDHF
kit (Lonza) according to the manufacturer's instructions. Equal concentrations
of the DMSO vehicle were used as controls for drug studies.

Kutys M.L, & Yamada K.M. (2014). An extracellular matrix-specific GEF-GAP interaction regulates Rho GTPase crosstalk for 3D collagen migration. Nature cell biology, 16(9), 909-917.

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HiChIP Experiments with HAEC and AoSMC

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HAEC (catalog no. CC-2535) and human AoSMC (catalog no. CC-2571) were obtained from Lonza. The HAEC were grown in endothelial growth medium (EGM medium), which is formulated by mixing the contents of the EGM SingleQuots kit (Lonza catalog no. CC-4133 containing bovine brain extract [BBE], ascorbic acid, hydrocortisone, epidermal growth factor [hEGF], fetal bovine serum [FBS], and gentamicin/amphotericin-B [GA] with EBM basal medium (Lonza catalog no. CC-3121). AoSMC were grown in SmGM-2 BulletKit (Lonza catalog no. CC-3182). The following growth supplements are added to a 500-mL bottle of smooth muscle cell basal medium: hEGF, 0.5 mL; insulin, 0.5 mL; hFGF-B, 1 mL; FBS, 25 mL; GA-1000, 0.5 mL. Both cell types were cultured at 37 °C in 5% CO₂. For cell culture expansion, trypsin/EDTA (ethylenediaminetetraacetic acid) was used for detachment of cells. Cells between passage 5 and 7 were used for the following HiChIP experiments.

Ma S., Chen X., Zhu X., Tsao P.S, & Wong W.H. (2021). Leveraging cell-type-specific regulatory networks to interpret genetic variants in abdominal aortic aneurysm. Proceedings of the National Academy of Sciences of the United States of America, 119(1), e2115601119.

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Oxidative Stress and Cell Viability

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HAECs, AoSMCs, and HUVECs were purchased from LONZA. HAEC and HUVEC were cultured in EGM-2 BulletKit (LONZA), while AoSMC was cultured in SmGM-2 BulletKit (LONZA), containing growth factors with 2% fetal bovine serum, at 37°C in a 5% CO₂ atmosphere. To examine the effects of oxidized human serum components, cells were grown on 48 well plates at a density of 4 x 10⁴ cells/ml (333 μl/well). After the cells were attached (16–18 h), they were treated with oxidized human serum components at 10% (33 μl/well) for 48 h. To determine cell viability, Premix WST-1 Cell Proliferation Assay System (TAKARA Bio Inc) was used. The cells were incubated with 10% WST-1 at 37°C for 2 h. The optical density of formazan was measured at 450 nm using a Multiskan Ascent plate reader (Thermo Electron).

Saito Y, & Noguchi N. (2016). Oxidized Lipoprotein as a Major Vessel Cell Proliferator in Oxidized Human Serum. PLoS ONE, 11(8), e0160530.

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PA-YK-NO Nanomat Viability Assay

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Human aortic smooth muscle cells (SMCs) (16,000 cells per each well; Lonza, Walkersville, MD) were seeded in the 24 well plate and cultured overnight with 1.5 ml of SMC growth medium (SmGM-2 BulletKit; Lonza) at normal cell culture conditions (37°C, 95% humidity, 5% CO₂). Then, varied concentrations of PA-YK-NO (0.2, 0.5, 1, and 3 wt.%) containing nanomatrix gels were loaded in the cell culture insert wells (3.0 μm pore size membrane, Corning, NC) and treated to the SMCs in the 24 well plate. The SMCs with the gels were cultured for 3 days under fetal bovine serum (FBS) deprived condition. After 3 days, the SMCs were stained by conducting a live/dead viability assay (Molecular Probes Inc., OR) and were quantified using microplate reader (Excitation/emission, live signal: 485 nm/528 nm and dead signal: 528 nm/617 nm).

Somarathna M., Hwang P.T., Millican R.C., Alexander G.C., Isayeva-Waldrop T., Sherwood J.A., Brott B.C., Falzon I., Northrup H., Shiu Y.T., Stubben C.J., Totenhagen J., Jun H.W, & Lee T. (2021). Nitric Oxide Releasing Nanomatrix Gel Treatment Inhibits Venous Intimal Hyperplasia and Improves Vascular Remodeling in a Rodent Arteriovenous Fistula. Biomaterials, 280, 121254.

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Primary Cell Culture Protocol

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Primary HUVECs were obtained from Lonza, Basel, Zwitserland (CC-2519) and ATCC, City of Manasses, Virginia, United States (PCS-100-010 and PCS-100-013) and were grown in EGM™-2 BulletKit™ (CC-3162, Lonza) according to the manufacturer’s instructions. Three different primary HASMCs were obtained from Lonza (CC-2571) and were grown in SmGM™-2 BulletKit™ (CC-3182, Lonza) according to the manufacturer’s instructions. For both HUVECs and HASMCs, growth media were renewed every 2 days and cells were sub-cultured using 0.05% Trypsin-EDTA (Invitrogen, Carlsbad, California, United States) when reaching a confluence of 70–80%.
HepG2 cells were grown in MEMα (12561056, Life Technologies, Carlsbad, California, United States) supplemented with 10% fetal bovine serum (Moregate, Hamilton, New Zealand). HEK293 cells were grown in DMEM (11995, Life Technologies) supplemented with 10% fetal bovine serum (Moregate, New Zealand). All cells were grown at 37°C, 5% CO₂.

Marsman J., Gimenez G., Day R.C., Horsfield J.A, & Jones G.T. (2019). A non-coding genetic variant associated with abdominal aortic aneurysm alters ERG gene regulation. Human Molecular Genetics, 29(4), 554-565.

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Culturing Human Coronary VSMC and EC

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Human coronary artery VSMCs and ECs (Lonza, Walkersville, MD) were cultured per the supplier’s instructions. VSMCs were maintained in Smgm-2 Bulletkit (Lonza, cat#CC4147 culture media; ECs were maintained in Ebm-2 Bulletkit (Lonza, cat#CC3156). Cells from passage 3 to 7 were used in experiments.

Yu D., Gernapudi R., Drucker C., Sarkar R., Ucuzian A, & Monahan T.S. (2019). The myristoylated alanine-rich C kinase substrate (MARCKS) differentially regulates kinase interacting with stathmin (KIS) in vascular smooth muscle and endothelial cells and potentiates intimal hyperplasia formation. Journal of vascular surgery, 70(6), 2021-2031.e1.

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Isolation and Culture of Pulmonary Arterial Smooth Muscle Cells

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Pulmonary arterial smooth muscle cells were isolated as described previously (Shimoda et al., 1998 ). Lungs were placed in cold HBSS and intrapulmonary arteries (PAs; 200–600 μm outer diameter) were isolated under a dissecting microscope and cleaned of adventitia and endothelium. The arteries were allowed to recover for at least 30 min on ice before being transferred to reduced-Ca²⁺ (20 μM CaCl₂) HBSS at room temperature for at least 20 min. The tissue was digested for 20–25 min at 37°C in reduced-Ca²⁺ HBSS containing collagenase (type I; 1,750 U/ml), papain (9.5 U/ml), bovine serum albumin (2 mg/ml) and dithiothreitol (1 mM). PASMCs were obtained by gentle trituration in Ca²⁺-free HBSS using a modified P1000 pipette tip and cultured in Smooth Muscle Cell Medium supplemented with SmGM™-2 bullet kit (Lonza). PASMC phenotype and purity were validated by assessing the presence of smooth muscle-specific actin (SMA) using immunofluorescence in cells incubated with anti-SMA (1:1,000, Sigma) primary antibody and counterstained with DAPI. PASMCs were used at passages 0–3.

Yun X., Philip N.M., Jiang H., Smith Z., Huetsch J.C., Damarla M., Suresh K, & Shimoda L.A. (2021). Upregulation of Aquaporin 1 Mediates Increased Migration and Proliferation in Pulmonary Vascular Cells From the Rat SU5416/Hypoxia Model of Pulmonary Hypertension. Frontiers in Physiology, 12, 763444.

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Cultivation of Aortic ECs and SMCs

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Primary human endothelial cells (ECs) and smooth muscles cells (SMCs) of aortic origin were purchased from Lonza (USA). ECs and SMCs were maintained in EGM-2 BulletKit (Lonza CC-3162) and SmGM-2 BulletKit (Lonza CC-3182) media, respectively.

Feaver R.E., Bowers M.S., Cole B.K., Hoang S., Lawson M.J., Taylor J., LaMoreaux B.D., Zhao L., Henke B.R., Johns B.A., Nyborg A.C., Wamhoff B.R, & Figler R.A. (2023). Human cardiovascular disease model predicts xanthine oxidase inhibitor cardiovascular risk. PLOS ONE, 18(9), e0291330.

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Vascular Cell Co-Culture Model for Studying MEJs

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Human umbilical vein endothelial cells (HUVECs), human coronary artery endothelial cells (HCAECs), and human coronary artery smooth muscle cells (HCASMCs) were cultured in EBM-2 supplemented with an EGM-2 bullet kit, EBM-2 supplemented with an EGM-2MV bullet kit, and SmGM-2 supplemented with a SmGM-2 bullet kit (Lonza, Basel, Switzerland), respectively. Human umbilical vein smooth muscle cells (HUVSMCs) and bovine aortic endothelial cells (BAECs) were obtained from Cell Applications, Inc. (San Diego, CA, USA) and cultured in growth medium according to the manufacturer’s instruction. All cells were cultured at 37°C and 5% CO₂. For a vascular cell co-culture model, ECs were co-cultured with vascular SMCs on a Transwell^(R) (24 mm) (Corning Inc., NY, USA) to allow a formation of MEJs, as previously described.^{11 (link),24 (link)} SMCs were first seeded on the lower part of the Transwell insert and cultured for 24 hours. In the same Transwell insert, ECs were cultured on the upper part of the insert and grown for an additional 24 hours. Following this initial period of growth, KLF2 or KLF4 expression was manipulated by adenoviral infection.

Sangwung P., Zhou G., Lu Y., Liao X., Wang B., Mutchler S.M., Miller M., Chance M.R., Straub A.C, & Jain M.K. (2017). Regulation of endothelial hemoglobin alpha expression by Kruppel-like factors. Vascular medicine (London, England), 22(5), 363-369.

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