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Continuous purification and culture of rat type 1 and type 2 alveolar epithelial cells by magnetic cell sorting

Chinese Journal of Traumatology 2022年第3期25卷 138-144页

作者： Di Liu Jian-Hui Sun Hua-Cai Zhang Jian-Xin Jiang Ling Zeng State Key Laboratory of Trauma Bums and Combined InjuriesDoping HospitalArmy Medical UniversityChongqing400042China Medical Center of Trauma and War Injury Daping HospitalArmy Medical UniversityChongqing400042China

Purpose:The incidence of acute lung injury(ALI)in severe trauma patients is 48%and the mortality rate following acute respiratory distress syndrome evolved from ALI is up to 68.5%.alveolar epithelial type 1 cells(AEC1 s)and type 2 cells(AEC2s)are the key cells in the repair of injured lungs as well as fetal lung ***,the purification and culture of AECls and AEC2s play an important role in the research of repair and regeneration of lung ***:Sprague-Dawley rats(3-4 weeks,120-150 g)were purchased for *** and DNase I were jointly used to digest lung tissue to obtain a single-cell suspension of whole lung cells,and then magnetic bead cell sorting was performed to isolate Tla positive cells as AECls from the single-cell suspension by using polyclonal rabbit anti-Tla(a specific AECls membrane protein)antibodies combined with anti-rabbit IgG ***,alveolar epithelial cell membrane marker protein EpCAM was designed as a key label to sort AEC2s from the remaining Tla-neg cells by another positive immunomagnetic selection using monoclonal mouse anti-EpCAM antibodies and anti-mouse IgG *** purity was identified by immunofluorescence staining and flow ***沾••The purity of AECls and AEC2s was 88.3%±3.8%and 92.6%±2.7%,*** cell growth was observed as follows:AECls stretched within the 12-16 h,but the cells proliferated slowly;while AEC2s began to stretch after 24 h and proliferated rapidly from the 2nd day and began to differentiate after 3 ***:AECls and AEC2s sorted by this method have high purity and good ***,our method provides a new approach for the isolation and culture of AECls and AEC2s as well as a new strategy for the research of lung repair and regeneration.

关键词： alveolar epithelial cells type 1 alveolar epithelial cells type 2 Three-dimensional culture Magnetic activated cell sorting

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