| The nerve regeneration chamber can solve the problem of short distance nerve regeneration, but the long distance nerve regeneration can't be solved by artificial repair technique except the autologous neural transplantation. The reasons that"artificial nerve"is not better than the nerve regeneration chamber with the development of tissue engineering of peripheral nervous system are the problems of seed cell. These problems such as cell senescence, degression of secreted function havn't be solved. Schwann cells (SCs) which are myelin-forming cells in peripheral nervous system are the most important seed cells. SCs can secrete many nerve nutrition factors which play a very important role during the regeneration of peripheral nerve. The secreted function of SCs can be influenced by multiple factors. Methylprednisolone (MP) can protect nerve cells and their prominence by the inhibition of lipid peroxidation.Objective:1. To find a good approach to obtain more population and more purification of SCs from new-born SD rats. Experimental studies on culture of SCs in vitro, while we observed SCs growth cases and the percentage of SCs purified.2. To explore the effects of methylprednisolone(MP) on the secreted function of cultured rats'SCs.Methods:1. To dissect the sciatic and brachial nerve of new-born SD rats. The nerve fascicles were then extracted under 10×microscope. The nerve was cut to be 1mm3 tissue pieces. Two enzymes were used to digest the sciatic nerve speciments. Cytarabine was used to be inhibition fibroblasts grow. Low density trypsin quickly digested to passage SCs. Then we adopted morphology observation, S-100 protein assessment.2. Different concentration of MP(10-3,10-4,10-6,10-8 mol/L) were administrated to the cell, while control group was given no drug. 24,48 and 72 hours after administration, the cells from different concentrations were measured with Reverse trancription-polymerase chain reaction (RT-PCR) methods respectively to measure the expression of NGF.Results:1. Through above-mentioned methods we got high purified Schwann cells of the second passage. The purity of SCs was 88%. The cell quantity was 3.128×107/ml. The morphology of most second passage cells was fusiform, the cell alignment liked whirlpool or palisade.2.After the measurement of the expression of NGF, we analyze the ratio of relatively density of NGF andβ-actin'straps with Lab works 4.60 and Spass 10.0. We got the statistic conclusion: the group which was given the MP with the concentration of 10-8 mol/L and was administrated the drug 72 hours expressed more NGF than other groups(p<0.05); while the groups which were given the MP with the concentration of 10-3 mol/L including the groups which were administrated the drug 24,48and72 hours expressed less NGF than other groups(p<0.05).Conclusions:1. We got SCs from SD rats'sciatic and brachial nerve. The method was convenient. This method provided cells resources for peripheral nerve tissue engineering. We got high quantity and high purified cells through above-mentioned methods.2. High concentration(10-3 mol/L) of MP prohibit secreted function of SCs, but long time and low dosage(72 hours,10-8 mol/L) MP promote secreted function of SCs.
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