人类胚胎嗅球嗅鞘细胞的分离培养及纯化方法研究

Objective To explore a convenient and pragmatic method to obtain sufficient high purity olfactory ensheathing cells (OECs) from human embryo olfactory bulb by using different attachment rates in harvested cells combined with the technique of applying adjusting serum at low concentration containing B-27 Supplement. Methods DMEM/F12 culture media supplemented with low concentration fetal bovine serum(2.5-5%) combined with adjusting serum including B-27 Supplement was used to culture olfactory ensheathing cells after differential velocity adherent intermittently every 48h. Observing the growth and proliferation changes of OECs in the whole culturing course. Moreover, the Purity of cultured OECs was identified with p75 nerve growth factor receptor (P75NGFR) fluorescent immunohistochemistry on the tenth day.Results This modified medium with B-27 supplement could promote the survival as well as the proliferation of OECs, which appeared to be dipolar or tripolar and processes formed a network in vitro. These cells were positive after using P75NGFR immunocytochemical staining, and the results showed theirs purity reached above 90% by this optimized method. Conclusions All of the above indicate that the optimized medium combined timely adjusting serum methods is effective in production of highly purified human embryo OECs.