摘要
目的改良原代肺泡巨噬细胞（alveolar macrophage,AM）培养技术,提高AM培养效果。方法SD大鼠,在体支气管灌洗肺获取AM,通过预先注射空气、14号注射器针头回收灌洗液、低速离心、种板前多次吹打等改良措施分离和培养AM。倒置显微镜连续观察AM的形态变化,台盼蓝拒染实验鉴定AM存活率,瑞氏-姬姆萨染色鉴定AM纯度。结果AM体外培养后40 min即见细胞贴壁;2～3天时,细胞形态多样,呈圆形、不规则形、梭形等形态,胞体变大,胞质丰富。台盼蓝拒染实验示AM存活率≥95%,瑞氏-姬姆萨染色示AM纯度≥95%。结论本实验改良了原代AM培养的技术方法,提高了原代AM培养效果。
Objective To modify the techniques for primary culture of highly purified the primary rat alveolar macrophages（AMs）.Methods AMs were isolated in vivo and cultured with such modified techniques as pre-injection of air and reclamation of brochoalveolar lavage fluid with 14-bugle syringe needle,low-speed centrifugation and multi-blowing before plating on 6-well culture plates.The morphological changes were observed under the inverted microscope.The cell survival rate was detected with trypan blue dye exclusion test,and cell purity was determined by Wright-Giemsa staining.Results In vitro,AMs adhered to plastic surface within 40 min;2-3 days later,cell morphology presented mixed shapes,i.e.,circular,irregular and spindle shapes.The trypan blue test showed a survival rate ≥95% of the AMs and Wright-Giemsa staining indicated a purity ≥95% of the AMs.Conclusion The modified techniques employed in our experiment had better results for the isolation and primary culture of AMs.
出处
《徐州医学院学报》
CAS
2010年第5期325-327,共3页
Acta Academiae Medicinae Xuzhou
关键词
肺泡巨噬细胞
细胞培养
技术改良
alveolar macrophage
cell culture
technique modification
