CAJ | 학술논문

背景:原代培养方法被广泛应用于正常和哮喘大鼠气道平滑肌细胞的培养,但少见用于慢性阻塞性肺疾病大鼠气道平滑肌细胞培养的报道. 目的:建立大鼠慢性阻塞性肺疾病模型,比较组织块法、酶消化法和改良组织块消化法原代培养模型大鼠气道平滑肌细胞生长情况的差异. 方法:将16只清洁级雄性健康SD大鼠随机分成2组,对照组正常饲养,慢性阻塞性肺疾病组用熏烟法建立大鼠慢性阻塞性肺疾病模型,显微镜下观察大鼠肺组织病理学特点.分别应用上述3种方法原代培养慢性阻塞性肺疾病大鼠气道平滑肌细胞,相差显微镜下观察细胞形态,用α-平滑肌肌动蛋白免疫组织化学染色鉴定细胞类型. 结果与结论:经病理学证实成功构建慢性阻塞性肺疾病大鼠模型.在倒置相差显微镜下见培养的细胞表现为典型的“谷-峰状”生长.经免疫细胞化学染色后,可见胞质呈棕色阳性反应,所培养的细胞有94%以上为气道平滑肌细胞.3种方法在耗时和细胞质量方面均无明显差别,但组织块法更经济、稳定可靠和简单.
배경:원대배양방법피엄범응용우정상화효천대서기도평활기세포적배양,단소견용우만성조새성폐질병대서기도평활기세포배양적보도. 목적:건립대서만성조새성폐질병모형,비교조직괴법、매소화법화개량조직괴소화법원대배양모형대서기도평활기세포생장정황적차이. 방법:장16지청길급웅성건강SD대서수궤분성2조,대조조정상사양,만성조새성폐질병조용훈연법건립대서만성조새성폐질병모형,현미경하관찰대서폐조직병이학특점.분별응용상술3충방법원대배양만성조새성폐질병대서기도평활기세포,상차현미경하관찰세포형태,용α-평활기기동단백면역조직화학염색감정세포류형. 결과여결론:경병이학증실성공구건만성조새성폐질병대서모형.재도치상차현미경하견배양적세포표현위전형적“곡-봉상”생장.경면역세포화학염색후,가견포질정종색양성반응,소배양적세포유94%이상위기도평활기세포.3충방법재모시화세포질량방면균무명현차별,단조직괴법경경제、은정가고화간단.
BACKGROUND:Methods of primary culture have been extensively used to culture airway smooth muscle cel s in normal or asthma rats, but the same way to culture airway smooth muscle cel s 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 cel s in rat chronic obstructive pulmonary disease models by three methods:attachment-block culture and enzymatic dispersion as wel 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 normal y. 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 cel s of rat chronic obstructive pulmonary disease model were cultured from primary generation using the three ways mentioned above, respectively. The morphology of the cultured cel s was observed under phase contrast microscope andα-smooth muscle actin immunostaining was used to identify the cel types. @@@@RESULTS AND CONCLUSION:The rat chronic obstructive pulmonary disease models were established successful y, which was proved by pathology. The cultured cel s demonstrated the typical“hil and val ey”appearance under phase contrast microscope. Afterα-smooth muscle actin immunostaining, brown positive reaction was observed in the cytoplasm. There were over 94%cultured cel s identified to be airway smooth muscle cel s. There was no significant difference of consuming time and quality of cel s 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 muscle cel s in rat chronic obstructive pulmonary disease models.