Aim: To explore a simple and pragmatic method to obtain sufficient olfactory ensheathing cells from human fetal by using different attachment rates in harvested cells with the combinating of the technique of using NT3 intermittently.
Methods: DMEM/F12 culture solution including 100 mL/L of fetal bovine serum or including NT3 was used to culture olfactory ensheathing cells intermittently every 48 h. The conditions and growth degree of OECs were observed, and P75 immunocytochemistry was used to estimate the purity of the cells.
Results: Human fetal OECs were positive after P75 immunocytochemistry. They appeared to be dipolar or tripolar cells and their processes formed a network in vitro. The purity of OECs in good conditions reached about 95% on 9 d and 83% on 12 d.
Conclusion: The method of using different attachment rates combined with the technique of using NT3 intermittently can culture and purify OECs simply and effectively.