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The purpose of the establishment of olfactory ensheathing cells (olfactory ensheathing cells, OECs) culture , identification and tracing methods , and preliminary to explore the feasibility of OECs cochlear implants . Method 1 , cell culture and purification : three days of neonatal Sprague-Dawley rat olfactory bulb sources OECs in primary culture , then by differential adhesion method anti- mitotic purified culture . Morphological characteristics and growth of the cells was observed at different time points in under an inverted microscope , using a low - affinity nerve growth factor receptor (NGFR-p75) by immunohistochemistry staining , and make the identification of cell purity . OECs 2 Animal experiments: The purified and cultured for 7 days with 5 - bromo- 2'-deoxy- uridine (5-bromo-2'-deoxyuridine, BrdUrd) OECs and incubated for 48 hours to mark . For adult guinea pigs incision , exposure , open bulla drilling nearly stapes artery in the basal turn of the cochlea , injecting OECs containing suspension . Cell transplantation after 1 day , take the guinea pig cochlea frozen sections , immunofluorescence staining , confocal laser distribution of transplanted cells under the microscope . 1 , purified and cultured OECs rapidly proliferating , up to nine days cell number , up to 85% purity . 2 , the BrdUrd and OECs common for 48 hours , its labeling rate can reach 90%. 3 , one day after OECs transplantation is mainly distributed in the guinea pig cochlea scala tympani . Conclusion 1 differential attachment wall law anti- mitotic method to obtain high purity of OECs . 2, BrdUrd successful marker of OECs is safe , effective , simple , marked the high rate of tracer method . Transplantation of OECs cochlear short-term survival in the scala tympani , and long-term outcomes needs further observation .
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