Primary culture of alveolar epithelial type II cells and its bionomic study.
To establish a better method of primary culture for alveolar epithelial type II cells (AEC II) and to study its bionomics, alveolar epithelial type II cells were isolated by digestion with trypsin and collagenase, which were then purified by plated into culture flask coated with rat immunoglobulin G. The purified AEC II were identified by alkaline phosphatase staining, electron microscopy, immunocytochemical staining of pulmonary surfactant protein A (SPA). The SPA expression and transfection characteristics were compared with those of A549 cell line. The results showed that AEC II could be isolated by digestion with trysin and collagenase and purified by adhesive purification by using IgG, with a yield of about 2-3 x 10(7), and a purity of about 75%-84%. Cells could be quickly identified with AKP staining. AEC II were different from A549 cell line in terms of SPA expression and transfection characteristics. It is concluded that adhesive purification with IgG can improve the purity of AEC II, and AKP staining is simple in cell identification. AEC II can not be completely replaced by A549 cells in some studies because the differences between them, such as SPA expression.
Shi
Xuemei
X
Research Institute of Respiratory Diseases, Department of Respiratory Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China. sxmxi2002@163.com
Zhang
Huilan
H
Xiong
Shengdao
S
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D004463:Ecology; D004847:Epithelial Cells / Q000166:cytology*; D007074:Immunoglobulin G / Q000494:pharmacology; D011650:Pulmonary Alveoli / Q000166:cytology*; D037662:Pulmonary Surfactant-Associated Protein A / Q000096:biosynthesis; D051381:Rats; D017208:Rats, Wistar
D007074:Immunoglobulin G; D037662:Pulmonary Surfactant-Associated Protein A