The Primary Research On Culturing, Identifying And Transplanting Into The Cochlea Of Olfactory Ensheathing Cells

Obiective: To establish the methods of culturing,identifying and tracing olfactory ensheathing cells (OECs)and to investigate the feasibility of transplanting the OECs into the cochlea initially.Methods:1,Cell culture and purification: The primary OECs were cultivated with the olfactory bulb of 3 days newborn Sprague-Dawley rats, purified with a differential attachment and anti-mitotic method. The morphological changes of the cultured OECs were observed under a phase contrast microscope at different culture time. We can identify them by their characteristic of expressing NGFRp75.The purity of the OECs were evaluated according to the percentage of NGFRp75 immunostaining cells.2,Animal experiment: Purified OECs were cultured for 7 days, then incubated with BrdUrd for 48 hours to mark OECs. The marked OECs were inoculated via a small hole drilled on the bony wall of scala tympani in the basal turn of the cochlea. At the first day after transplantation,the transplanted cells were examined under the confocal microscope to observe their distribution among cochlea.Results:1,In the process of the culture and purification,the maximum of the OECs appeared at the 9th day and their purifty was up to 85%.2,OECs were incubated with BrdUrd for 48 hours, more than 90% of them were labeled.3,At the first day after transplantation,OECs were detected in scala tympani of the cochlea in the guinea pigs. Conclusion:1,Using the cell culture technique of differential attachment with anti-mitotic,we can obtain high-purity OECs.2,Using BrdUrd technique , a safe,effective and simple method for marking cells,we can label OECs successfully.3,After transplanted into the cochlea, OECs could survive in the scala tympani for a short term. A further observation is needed to investigate the survival, biologic activity and fate of OECs in the cochlea in a longer duration.