Isolation, culture and identification of primary airway smooth muscle cells from mice
Received:January 25, 2016  
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DOI:10.3969/j.issn.1005-4847.2016.04.006
KeyWord:Airway smooth muscle cells, SMCs;Primary culture;Tissue section adherence;Immunofluorescence;MTT assay;Mice
              
AuthorInstitution
叶致豪 中南民族大学生命科学学院医学生物研究所, 武陵山区特色资源植物种质保护与利用 湖北省重点实验室, 武汉
许文豪 中南民族大学生命科学学院医学生物研究所, 武陵山区特色资源植物种质保护与利用 湖北省重点实验室, 武汉
刘庆华 中南民族大学生命科学学院医学生物研究所, 武陵山区特色资源植物种质保护与利用 湖北省重点实验室, 武汉
沈金花 中南民族大学生命科学学院医学生物研究所, 武陵山区特色资源植物种质保护与利用 湖北省重点实验室, 武汉
彭勇波 中南民族大学生命科学学院医学生物研究所, 武陵山区特色资源植物种质保护与利用 湖北省重点实验室, 武汉
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Abstract:
      Objective To develop a reliable method for the primary culture of airway smooth muscle cells (ASMCs) from mice by adherent tissue culture and to identify them by immunofluorescence microscopy. Methods Airway smooth muscle (ASM) tissue was isolated from BALB/c mice under dissecting microscope, and cut into 1 mm3 pieces. These tissue blocks were washed with PBS with 1% penicillin and streptomycin, and adhered on 6 cm culture dish with 5 mL culture media. The dishes were incubated at 37℃ in an incubator with 5% CO2. The obtained primary ASMCs were identified by immunofluorescence microscopy, and the cell proliferation was measured by MTT assay.Results We observed that obvious fusiform cells grew out from tissue blocks within 3 to 5 days. After 5 to 6 days, “hill-valley” structure was observed. After cell passage and purification, the immunofluorescence microscopy showed that the purity of isolated ASMCs reached up to 99%. The growth curve of ASMCs was constructed by MTT assay. Conclusions Obtained ASMCs from this simple and economical method show a preferable proliferation ability, density and purity, and can satisfy the need for cell biology research.
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