Human asm cells

Manufactured by Lonza

Sourced in Switzerland

Human ASM cells are a type of primary cell line derived from human airway smooth muscle tissue. They are designed for use in in vitro research applications.

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Lab products found in correlation

Pd98059, by Cell Signaling Technology (1 mentions) Irdye 680 lt conjugated polyclonal donkey anti mouse igg, by LI COR (1 mentions) Irdye 800 cw conjugated polyclonal goat anti rabbit igg, by LI COR (1 mentions) Polyclonal rabbit anti gapdh, by Merck Group (1 mentions) Polyclonal rabbit anti mlc, by Cell Signaling Technology (1 mentions) Odyssey infrared imaging system, by LI COR (1 mentions)

3 protocols using human asm cells

Murine lung fibroblast isolation and culture

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Primary murine lung fibroblasts were harvested following a modification of the protocol by Roman et al. and cultured with ±50 µg/ml nicotine, as previously described, and ±10–20 µM caffeic acid phenyl ester (CAPE, Sigma-Aldrich), an NFκB inhibitor, in complete serum free media for 24–72 hours after serum starvation overnight as noted in the figure legends [22] (link). Fibroblasts were also cultured ±10 µM of PD98059 (Cell Signaling, Danvers, MA), an ERK1/2 inhibitor, and ±10 µM c-Jun inhibitory peptide (Tocris Bioscience, Bristol, UK) to examine MAPK and c-Jun pathways [23] (link), [24] . Human ASM cells were purchased through Lonza (Portsmouth, NH) and cultured according to the manufacturer’s protocol. Primary murine ASM cells were harvested by incubating minced tracheal and bronchial tissue in DMEM F12/Ham’s media with 1 mg/ml dispase for 1 hour at 37°C. Tracheal and bronchial tissue pieces were then placed on plastic culture dishes and incubated at 37°C in a 5% CO₂ atmosphere in DMEM F12/Ham’s media until ASM cells migrated to form a monolayer on the plastic. Fibroblast and ASM primary cell harvests were verified using S100, von Willebrand factor and α-smooth muscle actin immunostaining.

Wongtrakool C., Grooms K., Bijli K.M., Crothers K., Fitzpatrick A.M, & Hart C.M. (2014). Nicotine Stimulates Nerve Growth Factor in Lung Fibroblasts through an NFκB-Dependent Mechanism. PLoS ONE, 9(10), e109602.

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Nicotine-Induced Smooth Muscle Cell Regulation

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After overnight serum starvation, human ASM cells (Lonza, Switzerland) ± nicotine (50 μg/ml, 72 h) and human ASM cells isolated from healthy and asthmatic donors were cultured in serum-free media. Primary antibodies employed included polyclonal rabbit anti-MLC (1:1000, Cell Signaling Technologies), polyclonal rabbit anti-phospho-MLC (1:500, Cell Signaling Technologies), and polyclonal rabbit anti-GAPDH (1:10,000, Sigma). Secondary antibodies used were IRDye 680LT conjugated polyclonal donkey anti-mouse IgG (1:20,000, LI-COR Biosciences) and IRDye 800CW conjugated polyclonal goat anti-rabbit IgG (1: 20,000, LI-COR Biosciences). Quantification of protein expression was performed by measuring integrated intensity using the Odyssey Infrared Imaging System (LI-COR Biosciences). Densitometry was used to quantify relative protein expression levels using Image J (National Institutes of Health). The band of the protein of interest was normalized to the appropriate loading control.

Galior K., Ma V.P., Liu Y., Su H., Baker N., Panettieri R J.r., Wongtrakool C, & Salaita K. (2018). Molecular Tension Probes to Investigate the Mechanopharmacology of Single Cells: A Step Towards Personalized Mechano-medicine. Advanced healthcare materials, 7(14), e1800069.

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Mouse ASM Cell Isolation and Culture

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Mouse ASM cell isolation and culture were performed as described elsewhere (32 (link)). Briefly, the trachea from C57BL/6J or B6.129S6-Casp7^tm1Flv/J (Caspase-7^−/−) mice was removed and transferred into Dulbecco’s modified Eagle’s medium (DMEM), supplemented with 10% fetal bovine serum (FBS), penicillin (100 U/ml), and streptomycin (100 μg/ml; culture medium). Connective tissue and airway epithelium were removed by firmly scraping the luminal surface. The trachea strips were cut into small pieces (~1 mm³) and cultured in culture medium at 37°C in 5% CO₂. ASM cells begin to migrate out of the fragments after 7 to 10 days. The cells were dissociated with 0.05% trypsin and subcultured in culture medium. Identification of mouse ASM cells was based on the morphology and expression of α-SMA. Mouse ASM cells of passage <6 were used in all the experiments. Human ASM cells were obtained from Lonza and grown in culture medium.

Shigemura M., Lecuona E., Angulo M., Homma T., Rodríguez D.A., Gonzalez-Gonzalez F.J., Welch L.C., Amarelle L., Kim S.J., Kaminski N., Budinger G.R., Solway J, & Sznajder J.I. (2018). Hypercapnia increases airway smooth muscle contractility via caspase-7-mediated miR-133a-RhoA signaling. Science translational medicine, 10(457), eaat1662.

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