Experimental Studies On Culture Of Schwann Cells In Vitro And Biocompatibility Between Schwann Cells And Biomaterials

Although the repair following injury of peripheral nerve had achieved a great progress in the recent years with the development of microsurgery, the results of the repair were still not satisfied. The preferred approach to repair peripheral nerve defect was to bridge the two ends of the injuried nerve with a segment of autologous nerve graft in clinic. Autologous nerve graft was the golden standard for the peripheral nerve defect repair. But this approach incured some donor site sense dysfunction and scarring. The limited availability of donor tissue represented a severe problem. With the development of tissue engineering, the lack of autologous nerve source possibly could be solved.Objective:1,To found a new approach to obtain more population and more purification of SCs(Schwann cells) from SD rats and promote SCs growing on different biomaterials. Experimental studied on culture of Schwann cells In Vitro, while we observed SCs growth cases and the percentage of SCs purified.2,We choosed the SCs which were high purity and high energy to transplant on different biomaterials(inaorporated chitosan conduits; sustained-release FK506 inaorporated chitosan conduits). We also observed the cell growth cases. We provided gist for tissue engineering bio-artifical nerve to repair the defect of peripheral nerve .Methods:1,SD rats'sciatic nerve was pre-degenerated for 10 days. Then SD rats'sciatic nerve was dissected. The nerve fascicles were then extractedunder 10×microscope. The nerve was cut to be 1mm3 tissue pieces. One enzyme was used to digest the sciatic nerve speciments. Geneticin was used to be inhibition fibroblasts grow. Low density enzyme quickly digested to passage Schwann cells. Then we adopted morphology observation, S-100 protein assessment.2,We transplanted the third generation cells which were digested by 0.125% enzyme to the two different biomaterials. Geneticin was also used to be inhibition fibroblasts grow. Meanwhile we observed cell growth cases. Then we compared the cell growth energy on different biomaterials. we also adopted morphology observation, S-100 protein assessment.Results:1,Through above-mentioned methods we got high purified Schwann cells. The purity of Schwann cells was 85%. The cell quautity was 2.764×107/ml. The morphology of most third passage cells was fusiform.2,The purity of Schwann cells which grew on sustained-release FK506 inaorporated chitosan conduits was 93%; on inaorporated chitosan conduits was 89%, simplicity culture of the fourth passage Schwann cells was 76%.3,The morphology of most fourth passage cells was fusiform. The cell growth energy was common. The size of OT was 0.1293. The cell alignment was not regular. The cell morphology which grew on inaorporated chitosan conduits was fusiform. The size of OT was 0.1568. The cell growth energy was better. The cell alignment was regular. The cell morphology which grew on sustained-release FK506 inaorporated chitosan conduits was fusiform. The size of OT was 0.1805. The cell growth energy was best of three groups. The cell alignment liked whirlpool or palisade.Conclusions:1,We got Schwann cells from SD rats'sciatic nerve. The method was convenient. This method provided cells resources for perpherial nerve tissue engineering. We got high quantity and high purifitied cells through above-mentioned methods.2,Biocompatibility between Schwann cells and biomaterial of chitosan conduits was good. FK506 effectively promoted Schwann cells growing.