Manual microdissection of ciliated cells from guinea pig's nasal epithelium / 中国组织工程研究

ABSTRACT

AIM: Respiratory tract cilia are difficult to harvest and observe due to weak transparence, which results from their location in thick mucosa, so people always try to obtain ciliated cells with many methods. This study was designed to apply manual microdissection to isolate ciliated cells from guinea pig's nasal epithelium and offer pure cells to further research. METHODS: The experiment was carried out at the Institute of Otolaryngology Head and Neck Surgery in Chinese PLA General Hospital from March of November in 2007. Twenty healthy white guinea pigs were recruited in this study. The ciliated cells layer was isolated from submucous layer by instruments for microsurgery under the light microscope. The isolated mucosa and untreated mucosa layer were stained with hematoxylin-eosin in order to verify the isolated layer is mucous ciliated cells layer. The protease-treated ciliated cells layer was viewed under low magnification microscope, to count number of single ciliated cells and impure cells, as well as size of ciliated cells group. The morphology of ciliated cells was observed under high magnification microscope. RESULTS: Result of hematoxylin-eosin stain confirmed that the cell layer by manual microdissection was ciliated cells and basal cell layer, which were thin without submucous layer. Under low magnification microscope after manual microdissection, the density of isolated ciliated cells was larger, number of single ciliated cells was more and size of ciliated cells group was smaller. There were about 7 ciliated cells in a cell group, and no other cells were observed. Under high magnification microscope, the isolated ciliated cells by manual microdissection were smooth, good in light refraction, and active for cilia beat. The survival time was prolonged obviously, still 50% cells were alive 24 hours later. Compared with conventional isolation technique, manual microdissection was significantly predominant in the purity and activity of cells (P