Survival and number of olfactory ensheathing cells transplanted in contused spinal cord of rats / 中华创伤杂志(英文版)

<p><b>OBJECTIVE</b>To observe the survival and the number of olfactory ensheathing cells (OECs) transplanted in the contused spinal cord, so as to provide a basis for further studying the biological action of OECs.</p><p><b>METHODS</b>The rat spinal cords were contused with NYU-impactor II at T10 level by dropping a 10 g rod from a height of 25 mm. At the 1st week after injury, OECs isolated freshly from green fluorecense protein (GFP) of the rats were transplanted into the spinal cord at injured site and other two sites 1 mm apart from the caudal and rostral ends with the OECs number of 30000/μl x 3 = 90000. The survival and the number of OECs were qualitatively and semi-quantitatively observed under the fluorescense microscope from 1 week to 13 weeks after transplantation. The motor function of the cord was evaluated with BBB score.</p><p><b>RESULTS</b>GFP-OECs could survive at least for 13 weeks within the contused spinal cord. Their arrangement was from tight to loose and their number was decreased from 1 week to 13 weeks after injury. The average number of GFP-OECs was 536 at the 1st week, which was less than 1% of the number as compared with original transplantation. After then, the number of GFP-OECs was continually decreased, but the most obvious decrease was found during 1 week to 2 weeks. The extent of decrease at other time points was relatively mild. In contrast to the cell number, motor function of the cord was gradually recovered after transplantation.</p><p><b>CONCLUSIONS</b>The survival and the number of GFP-OECs are different between the animals and are affected by the pathological reaction of the host cord. Also it is related to the motor function recovery of the contused cord.</p>

Culture and morphologic characteristics of olfactory ensheathing cells isolated from young adult GFP rat / 中华神经医学杂志

Objective To establish methods for isolating and culturing the olfactory ensheathing cells (OECs) from young adult GIT rat to lay the foundation for repair of the spinal cord injury.Methods The OECs were dissociated from the first outer layer of the olfactory bulb of the young adult GFP rat (2.5 months old) under anatomical microscope; enzymatic digestion was performed on these cells and then they were inoculated into DMEM/F12 with 20% fetal calf serum. The morphology of OECs was regularly observed and photographied under light and electronic microscope. On the 10th culture day, the OECs were identified by the immunocytochemistry staining of S-100 and NGFRp75 and the purity (the positive rate) was calculated. Results The OECs showed strong green fluorescence under fluorescent microscope and presented morphological types of bipolar and bearing multipolar. More than 95% of the cultured cells were identified to be OECs, which expressed S-100 and NGFR p75. The cell structure revealed by electron microscope was much accordance with that by light microscope. Conclusion This method is easy to perform and high purity of GFP-OECs can be harvested with the same morphological characteristics and biological activity as general OECs. Therefore, OECs derived from the GFP transgenic rat can be effective tool cells, and widely used in studying the role of OECs in the repair of spinal cord injury