Culture and purification of human fetal olfactory bulb ensheathing cells*

ObjectiveTo obtain high purity of human fetal olfactory bulb ensheathing cells (OB-hOECs) in vitro and to develop a simple and effective method for primary culture of OB-hOECs.MethodsOB-hOECs were cultured based on the differential rates of attachment of the various harvested cell types. Then the method was combined with arabinoside cytosine (Ara-C) inhibition, serum-free starvation or intermittent neurotrophin 3 (NTS) nutrition method to observe cell states in different cultural environments. The purity of OB-hOECs was assessed with immunocytochemical analysis.ResultsOB-hOECs appeared bipolar and tripolar shape, with slender processes forming network. The purity of OECs reached 88% with the selective attachment method on day 6, and then fibroblast proliferated quickly and reduced the purity. When combined with the starvation method, the purity of OECs was 91% on day 6 and 86% on day 9, however, OECs were in a poor state. While combined with the NTS method, the purity reached 95% on day 9 and 83% on day 12, respectively. The cells still remained in a good state.ConclusionA combination of selective attachment and intermittent NTS nutrition is an effective method to obtain OECs with higher purity and quality.