四川大学学报（医学版）2017，Vol.48Issue(2)：282-285,308,5.
新生大鼠Ⅰ型肺泡上皮细胞原代培养方法的改进
The Improvement in the Primary Culture of Alveolar Epithelial Type Ⅰ of Rats
蒋玲 1胡远东 1徐菲菲 1王廷华1
作者信息
- 1. 四川大学华西医院麻醉科,成都610041
- 折叠
摘要
Abstract
Objective To improve the method of culturing and obtain purified alveolar epithelial type Ⅰ (AT Ⅰ) cells of SD rats.Methods 1 d newborn SD rats were applied for cell culture and brains were decapitated for lung tissues obtaining after respiratory cessation.Collagenase Ⅰ and DNase Ⅰ were used to digest and isolate cells.Then,cells were put into the plate coated with type Ⅰ rat tail collagen and different adherence was used to purify cells.Meanwhile,culture medium replacement was conducted after 48 h.The growth status was observed under an inverted microscope.Immunofluorescence including specific marker AQP5,SPC,BSI,Vimentin were used to identify cells.Results 2 d after incubation,the cells began to adhere to plate.At day 4 and 6,cells began to proliferate and exhibited a shape of spindle,cube and polygon.8 d after incubation,the character of proliferation reached the highest and the cell viability was (87+8)％ and purification was (87±5)％.Conclusion By modifying the methods of tissues harvest,isolation and culture,we can obtain AT Ⅰ cells with high yield,good growth state and super purity.关键词
Ⅰ型肺泡上皮细胞/原代培养/鉴定Key words
Alveolar epithelial type Ⅰ/Primary culture/Identification引用本文复制引用
蒋玲,胡远东,徐菲菲,王廷华..新生大鼠Ⅰ型肺泡上皮细胞原代培养方法的改进[J].四川大学学报（医学版）,2017,48(2):282-285,308,5.
