Analysis Of Different Proteomics Between Human Umbilical Cord Blood Mesenchymal Stem Cells Derived Schwann-like Cells And Fetal Sciatic Nerve’s Schwann Cells

Background: The clinical treatment of peripheral nerve injury defect is the world’sproblems, nerve autograft is the gold method, but for the area will cause nerve damage.By tissue-engineered artificial nerve repair peripheral nerve defects is a hot clinicalstudies for treatment. ANN seed cells for Schwann cells (Schwann cells, SCs), its mainsource of autologous SCs, allogeneic SCs, as well as from mesenchymal stem cells(MSCs) induced to differentiate from, and MSCs induced SCs come to class a widerange of its source, strong activity favored more by scholars. Human umbilical cordblood-derived mesenchymal stem cells (Human Umbilical Cord Blood MesenchymalStem Cells, HUCBMSCs) as an alternative to the first application of human bonemarrow mesenchymal source of stem cells (Mesenchymal stem cells, MSCs) in vitroculture has proved it has SCs can differentiate the class, but concrete and normal SCsWhat similarities and differences to be further studied.Objective: This study aims to establish human embryonic SCs of sciatic isolation,culture, purification and identification system, combined with our group pastHUCBMSCs vitro culture and experience, the class SCs HUCBMSCs, HUCBMSCsafter in vitro and human SCs three cell the analysis of proteomics, protein spots tofind.Methods: The research team past HUCBMSCs culture and in vitro methods and classesto get HUCBMSCs SCs; take the drug induction of1012to16Zhouren embryonicsciatic nerve membrane peel, cut into pieces, digestion, centrifuge, training, to becovered with bottles after the bottom of group A with low enzyme digestion anddifferential adhesion and L-Valine conditioned medium was purified SCs; group Benzymatic digestion combined with low differential adhesion purification purification;group C L-Valine purified conditioned medium France; D treatment group did not useany purification. Purity of cells in each group, select a group of the highest purity forthe next step experiment Schwann cells under a microscope the unique morphology statistics. Select three cells, total protein was extracted after two-dimensional gelelectrophoresis protein utilization and silver staining, were analyzed and comparedusing ImageMaster software, select HUCBMSCs expression compared with SCs andHUCBMSCs between the low point during induction increased protein spots massspectrometry match point, and select the appropriate protein immunoblot verified.Results:1) low enzyme digestion and differential adhesion after L-Valine conditionedmedium purity of purified SCs5d reached99.1%.2) obtain a good resolution andrepeatability are two-dimensional gel electrophoresis patterns of protein expression ineach gel point is3500or so, the average match rate was98%,95%,97%.3) beforeinduction HUCBMSCs and protein differences SCs have290points after inductionclass SCs and SCs only161protein spots, indicating HUCBMSCs closer afterinduction SCs in the composition of proteins.4) screened41proteins matching points,including25protein-point difference over three times, taking it25proteins by massspectrometry analysis of the final total of20points are successfully identified,belonging cytoskeletal proteins, cell regulatory proteins, energy metabolism andprotein protease protein, which contains several recognized SCs labeled protein.5)Select the three kinds of proteins in20protein spots in immunoblotting validationverification results are basically consistent with the results of two-dimensionalelectrophoresis and mass spectrometry.Conclusion:1.Through low enzymatic digestion, differential adherence and L-Valineconditioned medium combining culture and purification methods SCs SCs shorter time,higher purity, can provide for the next nerve cell source for tissue engineering.2. HUCBMSCs closer to normal after the induction of SCs in the composition of theprotein.3.This experiment successfully identified20protein spots belonging cytoskeletalproteins, cell regulatory protein, energy metabolism and protein protease protein,protein content of these changes are due to the inducer stimulate cells to produce themajority of the nerves differentiation and growth plays an important role, which alsoincludes several recognized SCs labeled protein.4These differences in protein looking for better optimization of future programs as wellas in vitro guidance of charge between mesenchymal stem cells in vitro inducedorientation provide a reliable basis.