An improved method for guinea pig airway smooth muscle cell culture and the effect of SPFF on intracellular calcium

Liu,Jian; Zhang,Yuyang; Li,Qian; Zhuang,Quankun; Zhu,Xiaojie; Pan,Li; Cheng,Maosheng

doi:10.3892/mmr.2014.2385

September-2014 Volume 10 Issue 3

Full Size Image

Journals

International Journal of Molecular Medicine

International Journal of Molecular Medicine is an international journal devoted to molecular mechanisms of human disease.

International Journal of Oncology

International Journal of Oncology is an international journal devoted to oncology research and cancer treatment.

Molecular Medicine Reports

Covers molecular medicine topics such as pharmacology, pathology, genetics, neuroscience, infectious diseases, molecular cardiology, and molecular surgery.

Oncology Reports

Oncology Reports is an international journal devoted to fundamental and applied research in Oncology.

Experimental and Therapeutic Medicine

Experimental and Therapeutic Medicine is an international journal devoted to laboratory and clinical medicine.

Oncology Letters

Oncology Letters is an international journal devoted to Experimental and Clinical Oncology.

Biomedical Reports

Explores a wide range of biological and medical fields, including pharmacology, genetics, microbiology, neuroscience, and molecular cardiology.

Molecular and Clinical Oncology

International journal addressing all aspects of oncology research, from tumorigenesis and oncogenes to chemotherapy and metastasis.

World Academy of Sciences Journal

Multidisciplinary open-access journal spanning biochemistry, genetics, neuroscience, environmental health, and synthetic biology.

International Journal of Functional Nutrition

Open-access journal combining biochemistry, pharmacology, immunology, and genetics to advance health through functional nutrition.

International Journal of Epigenetics

Publishes open-access research on using epigenetics to advance understanding and treatment of human disease.

Medicine International

An International Open Access Journal Devoted to General Medicine.

September-2014 Volume 10 Issue 3

Full Size Image

Article

An improved method for guinea pig airway smooth muscle cell culture and the effect of SPFF on intracellular calcium

Authors:
- Jian Liu
- Yuyang Zhang
- Qian Li
- Quankun Zhuang
- Xiaojie Zhu
- Li Pan
- Maosheng Cheng
View Affiliations / Copyright

Affiliations: School of Life Science and Biopharmaceutics, Shenyang Pharmaceutical University, Shenyang, Liaoning 110016, P.R. China, School of Pharmaceutical Engineering, Shenyang Pharmaceutical University, Shenyang, Liaoning 110016, P.R. China
Pages: 1309-1314
|
Published online on: July 15, 2014

https://doi.org/10.3892/mmr.2014.2385
Expand metrics +

Abstract

The aim of the present study was to establish an improved method for in vitro guinea pig airway smooth muscle (ASM) cell culture and to evaluate the effect of 2-(4-amino-3-chloro-5-triﬂuomethyl-phenyl)-2-tert-butylamino-ethanol hydrochloride (SPFF), a novel β2-adrenoceptor agonist, on the release of intracellular calcium in cells. A procedure for the efficient isolation, culture, passage and characterization of the cells was described. Primary ASM cells of guinea pigs were cultured by modified tissue cultivation. The cells were identified by their morphological characteristics and immunocytochemistry. The relative inhibition of the release of intracellular calcium by drugs in the cells was measured by fluorometric quantification with fluorochrome Fura-2/AM. The results were as follows: a) The ASM cells of the guinea pigs were successfully cultured and subcultured by using our improved method and typical peak-valley characteristics were observed under the phase contrast microscope; b) data from immunocytochemical staining with specific α-smooth muscle actin (α-SMA) demonstrated that the cells were ASM cells; c) the growth characteristics and cell viability demonstrated that the cells were in good condition and were able to be applied in the follow-up studies; d) the inhibitory effect of SPFF on the release of intracellular calcium was concentration‑dependent when compared with the control and e) the potential mechanisms of SPFF on the inhibition of intracellular calcium may be independent of the ryanodine receptor, but may be closely associated with the inositol 1,4,5-trisphosphate receptor.

View Figures

View References

1	Camoretti-Mercado B: Targeting the airway smooth muscle for asthma treatment. Transl Res. 154:165–174. 2009. View Article : Google Scholar : PubMed/NCBI
2	Woodruff PG: Gene expression in asthmatic airway smooth muscle. Proc Am Thorac Soc. 5:113–118. 2008. View Article : Google Scholar : PubMed/NCBI
3	Liu QH, Zheng YM and Wang YX: Two distinct signaling pathways for regulation of spontaneous local Ca2+ release by phospholipase C in airway smooth muscle cells. Pflugers Arch. 453:531–541. 2007.PubMed/NCBI
4	Ethier MF and Madison JM: IL-4 inhibits calcium transients in bovine trachealis cells by a ryanodine receptor-dependent mechanism. FASEB J. 20:154–156. 2006.PubMed/NCBI
5	Pelaia G, Renda T, Gallelli L, et al: Molecular mechanisms underlying airway smooth muscle contraction and proliferation: implications for asthma. Respir Med. 102:1173–1181. 2008. View Article : Google Scholar : PubMed/NCBI
6	Govindaraju V, Michoud MC, Al-Chalabi M, Ferraro P, Powell WS and Martin JG: Interleukin-8: novel roles in human airway smooth muscle cell contraction and migration. Am J Physiol Cell Physiol. 291:C957–C965. 2006. View Article : Google Scholar : PubMed/NCBI
7	Gan LL, Wang MW, Cheng MS and Pan L: Trachea relaxing effects and beta2-selectivity of SPFF, a newly developed bronchodilating agent, in guinea pigs and rabbits. Biol Pharm Bull. 26:323–328. 2003. View Article : Google Scholar : PubMed/NCBI
8	Mosmann T: Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 65:55–63. 1983. View Article : Google Scholar : PubMed/NCBI
9	Hao Z, Zhang Y, Pan L, et al: Comparison of enantiomers of SPFF, a novel beta2-Adrenoceptor agonist, in bronchodilating effect in guinea pigs. Biol Pharm Bull. 31:866–872. 2008. View Article : Google Scholar : PubMed/NCBI
10	Madison JM and Ethier MF: Interleukin-4 rapidly inhibits calcium transients in response to carbachol in bovine airway smooth muscle cells. Am J Respir Cell Mol Biol. 25:239–244. 2001. View Article : Google Scholar : PubMed/NCBI
11	Matsumoto H, Hirata Y, Otsuka K, et al: Interleukin-13 enhanced Ca2+ oscillations in airway smooth muscle cells. Cytokine. 57:19–24. 2012. View Article : Google Scholar : PubMed/NCBI

Journals

International Journal of Molecular Medicine

International Journal of Oncology

Molecular Medicine Reports

Oncology Reports

Experimental and Therapeutic Medicine

Oncology Letters

Biomedical Reports

Molecular and Clinical Oncology

World Academy of Sciences Journal

International Journal of Functional Nutrition

International Journal of Epigenetics

Medicine International

An improved method for guinea pig airway smooth muscle cell culture and the effect of SPFF on intracellular calcium

This article is mentioned in:

Abstract

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Related Articles