Morphometric Analysis And Identification Of Epithelial Stem Cell Antigens In The Conducting Airway Of Porcine

The lung is unique for its gas-exchanging function, however, Airway epithelium is exposed directly to microrganisms, substances and pollutants in the air. Meanwhile lung epithelial itself is constantly engaged in self-repair to ensure normal lung function. It has been demonstrated that airway epithelial progenitor/stem cells play the crucial role In the process of injury and repair. Therefore, the isolation and identification Airway epithelial stem/progenitor cell,(EpiSPC) are meaningful for comprehend the function of airway epithelium, also for the treatment of pulmonary disease. However it’s difficult to isolate and identifie human airway epithelium cells due to the few resources of human lung tissue, while the proportion of stem cells is quite rare in human lung. The use of animal models could help a lot in-establishing Isolation and Identification method of lung epithelial stem cells, it promotes the study of human lung stem cells.Porcine is frequently used Among the models animals because it’s lung is similar with human’s in size, anatomical structures and biological characteristics of epitheliun cells. Therefore, to study Porcine stem cell surface markers are meaningful for isolate and identify Porcine tracheal epithelial ancestors/stem cells, even meaningful for future study of lung signaling pathway controling, it is functions for human lung stem cell biology when providing a reliable theoretical basis and experimental technology platforms in maintaining lung homeostasis, studing lung disease occurrence mechanisms and promoting lung tissue engineering research. To this end, we have carried out research work related to pig tracheal epithelium, and obtained the following results. These results lay a foundation for understanding Morphometric and sorting identification of stem cell in the large airway of porcine, which provide useful information for further studies of the regulatory mechanism of lung stem cells in a porcine model.1、The morphological characteristics of cell types and compositions in porcine airway epithelia will be analyzed in this study.The epithelial cell types and morphometry of epithelia in the large airway of porcine were investigated by using HE (Hematoxylin Eosin) and PAS (Periodic Acid-Schiff), as well as immunohistochemistry staining (IHC) using antibodies against various epithelial cell markers in the present study, the distinct compositions of epithelial cell types among different segments in the large airway of porcine, in which the majority of cell types were basal cells, ciliated cell, goblet cells and Clara cells. Such a morphometric characterization is similar to that found in humans, but differ from that seem in the murine airway. Together with findings from others, these results strongly evidenced that the porcine lung may be a better model for lung diseases in comparison with that of murine lung.2、The majority of cell types composed in the distal airway of porcine are basal cells, ciliated cell, goblet cells and Clara cells. The expressions of stem cell specific antigens and their specific antibodies were evaluated by an immunofluorescent staining in the present study. The distal airway epithelial cells were evaluated for the expression of markers of epithelial cell types and stem cells by immunofluorescent staining on frozen sections or cytoslides using distinct specific antibodies. Immunofluorescent analysis showed no detectable expression of specific antigens for airway epithelial stem cells on the airway frozen sections when the antibodies against10potential stem cell markers (such as CD117, Sea-1、Sox2、CD49f and SSEA-1) were utilized. However, the CD49f and CD117were detected to co-express with Keratin14in a subset of epithelial cells on cytoslides. Only infrequent stem cells reside in the epithelium for detection at a stead state of lung. Upon the injury (such as dissociation of epithelial cells), the stem cells is able to undergo proliferation for injure repair, which in turn to elevate the expression of specific antigens of stem cells. Thus, it is a practicable approach using a combination of Keratin14and CD49f or CD117for stem cell sorting as double stem cells, by which the resulting subpopulation(s) of potential stem cells could be used for studies of cell biology and functions.