Culture and purification of human fetal olfactory ensheathing cells using different attachment rates combined with intermittent NT3 nutrition1

ObjectiveTo explore a simple and pragmatic method to obtain sufficient olfactory ensheathing cells from human fetus by selective attachment of harvested cells combined with intermittent NT3 nutrition.MethodsDMEM/F12 culture solution including 10% fetal bovine serum or NT3 was used to culture olfactory ensheathing cells intermittently every 48 h. The cell state and growth rates of OECs were observed, and P75 staining was used to estimate the purity of the cells.ResultsHuman fetal OECs were positive with P75 immunocytochemical staining. OECs in dipolar or tripolar shape formed networks by their processes in vitro. The purity of OECs in “good state” was about 95% at 9 d and 83% on 12 d, respectively.ConclusionThe method of using different attachment rates combined with intermittent NT3 addition is a simple and effective way to culture and purify OECs.