To explore a method for isolating, culturing, and characterizing the olfactory ensheathing cells from the human middle turbinate tissues obtained during surgery.

The mucosa tissues of the middle turbinate was collected during surgery, and the smaller tissue blocks were obtained by two-step digestion and stripping of the mucosal epithelial tissues. The small tissue blocks were then cultured and pressure screening were performed during the culture for isolating the bipolar and multipolar cells. These cells were further identified by a staining with an assay of immunofluorescence. The ratio of epithelial-like cells, S100β as well as p75 positive cells was expressed as ±s; and compared by independent sample t-test.

Comparing the ratio of the epithelial-like cells in the primary cells cultured with or without the epithelial layer, the ratio of epithelial-like cells in the group with the epithelial layer was (92.23±3.93) %, which was higher than that in the group without the epithelial layer (77.63±2.97) % (t = 5.129, P = 0.007) . The primary cells were cultured by the stripping epithelial layer. The cells subjected to culture and pressure screening were bipolar or multipolar-like, which conformed to the morphological characteristics of olfactory ensheathing cells. Immunofluorescence staining confirmed that (8.1±1.7) % cells showed S100β positive and less than 5% cells showed p75 positive in the total population of the cells.

By using a two-step digestion method and a pressure-screening method, the human middle turbinate olfactory ensheathing cells were successfully obtained from the human nasal mucosa tissue, and the cell separation and culture period was significantly shortened compared with the conventional method.