| Primary Culture of Alveolar Epithelial Type Ⅱ Cells and Its Bionomic Study |
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| 引用本文： | 史雪梅,张惠兰,熊盛道,甄国华,熊维宁,张珍祥,徐永健,胡琼洁,赵建平,倪望.Primary Culture of Alveolar Epithelial Type Ⅱ Cells and Its Bionomic Study[J].华中科技大学学报(医学英德文版),2007,27(6):653-656. |
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| 作者姓名： | 史雪梅 张惠兰 熊盛道 甄国华 熊维宁 张珍祥 徐永健 胡琼洁 赵建平 倪望 |
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| 作者单位： | Research Institute of Respiratory Diseases Department of Respiratory Diseases Tongji Hospital Tongji Medical College Huazhong University of Science and Technology，Research Institute of Respiratory Diseases Department of Respiratory Diseases Tongji Hospital Tongji Medical College Huazhong University of Science and Technology，Research Institute of Respiratory Diseases Department of Respiratory Diseases Tongji Hospital Tongji Medical College Huazhong University of Science and Technology，Research Institute of Respiratory Diseases Department of Respiratory Diseases Tongji Hospital Tongji Medical College Huazhong University of Science and Technology，Research Institute of Respiratory Diseases Department of Respiratory Diseases Tongji Hospital Tongji Medical College Huazhong University of Science and Technology，Research Institute of Respiratory Diseases Department of Respiratory Diseases Tongji Hospital Tongji Medical College Huazhong University of Science and Technology，Research Institute of Respiratory Diseases Department of Respiratory Diseases Tongji Hospital Tongji Medical College Huazhong University of Science and Technology，Research Institute of Respiratory Diseases Department of Respiratory Diseases Tongji Hospital Tongji Medical College Huazhong University of Science and Technology，Research Institute of Respiratory Diseases Department of Respiratory Diseases Tongji Hospital Tongji Medical College Huazhong University of Science and Technology，Research Institute of Respiratory Diseases Department of Respiratory Diseases Tongji Hospital Tongji Medical College Huazhong University of Science and Technology，Wuhan 430030 China，Wuhan 430030 China，Wuhan 430030 China，Wuhan 430030 China，Wuhan 430030 China，Wuhan 430030 China，Wuhan 430030 China，Wuhan 430030 China，Wuhan 430030 China，Wuhan 430030 China |
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| 基金项目： | This project was supported by grants from the National Natural Sciences Foundation of China （No, 30500224 and No. 30400193）. |
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| 摘 要： | 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 tryp- sin and collagenase, which were then purified by plated into culture flask coated with rat immu- noglobulin G. The purified AEC Ⅱ were identified by alkaline phosphatase staining, electron mi- croscopy, immunocytochemical staining of pulmonary surfactant protein A (SPA). The SPA expres- sion 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 pu- rification by using IgG, with a yield of about 2–3×107, and a purity of about 75%–84 %. Cells could be quickly identified with AKP staining. AECⅡ 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 AECⅡ, and AKP staining is simple in cell identification. AECⅡ can not be completely replaced by A549 cells in some studies because the differences between them, such as SPA expression.
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| 关 键 词： | alveolar epithelial type Ⅱ cells primary culture bionomics |
| 收稿时间： | 2007-09-23 |
Primary culture of alveolar epithelial type II cells and its bionomic study |
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| Shi Xuemei,Zhang Huilan,Xiong Shengdao,Zhen Guohua,Xiong Weining,Zhang Zhenxiang,Xu Yongjian,Hu Qiongjie,Zhao Jianping,Ni Wang.Primary culture of alveolar epithelial type II cells and its bionomic study[J].Journal of Zuazhong University of Science and Technology: Medical Edition,2007,27(6):653-656. |
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| Authors: | Shi Xuemei Zhang Huilan Xiong Shengdao Zhen Guohua Xiong Weining Zhang Zhenxiang Xu Yongjian Hu Qiongjie Zhao Jianping Ni Wang |
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| Institution: | 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 |
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| Abstract: | 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. |
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