Abstract:

BACKGROUND: Methods of primary culture have been extensively used to culture airway smooth muscle cells in normal or asthma rats, but the same way to culture airway smooth muscle cells in chronic obstructive pulmonary disease rats has rarely been reported.
OBJECTIVE: To establish the rat chronic obstructive pulmonary disease model, and to compare the different growth conditions of airway smooth muscle cells in rat chronic obstructive pulmonary disease models by three methods: attachment-block culture and enzymatic dispersion as well as improved tissue-piece digestion inoculation.
METHODS: Sixteen healthy male clean-grade Sprague-Dawley rats were randomly divided into chronic obstructive pulmonary disease group and control group with eight rats in each group. Rats in the control group were fed normally. The rat chronic obstructive pulmonary disease models were established in the chronic obstructive pulmonary disease group by cigarette smoke exposure. The pathologic characteristics of lung tissues of the rat models were observed under microscope. The airway smooth muscle cells of rat chronic obstructive pulmonary disease model were cultured from primary generation using the three ways mentioned above, respectively. The morphology of the cultured cells was observed under phase contrast microscope and α-smooth muscle actin immunostaining was used to identify the cell types.
RESULTS AND CONCLUSION: The rat chronic obstructive pulmonary disease models were established successfully, which was proved by pathology. The cultured cells demonstrated the typical “hill and valley” appearance under phase contrast microscope. After α-smooth muscle actin immunostaining, brown positive reaction was observed in the cytoplasm. There were over 94% cultured cells identified to be airway smooth muscle cells. There was no significant difference of consuming time and quality of cells in these three ways. Compared with the other two methods, attachment-block culture was a more economical and stable and less complicated way to culture airway smooth musclecells in rat chronic obstructive pulmonary disease models.