| ObjectiveIn vitro culture and identify isolated mesenchymal stem cells of human umbilical cord andSchwann cells of rats sciatic nerve, construct the chitosan composite scaffolds withChitosan-NT3cells, and implant the scaffolds into an established rat model of spinal cord injury,detect chitosan composite scaffolds for repair effect of rat spinal cord transection injury bysetting the control experiments and related indexes.MethodsIsolation, culture of human umbilical cord mesenchymal stem cells by enzyme digestion, andcultured, amplified. The use of cell morphology observed by optical microscope, to observe thecell growth curve identified by immunophenotype by flow cytometry. SD rat sciatic nerveSchwann cells cultured in isolation, and passage, were identified by immunohistochemicalmethod, and optical microscope was used to observe the cell morphology. Preparation ofchitosan (Chitosan-NT3scaffold materials provided by the Jinan University Institute ofTechnology), human umbilical cord mesenchymal stem cells, rat Schwann cells for scaffoldcomposite material implanted into rat model compound by setting the corresponding controlgroup, to observe its effect on repair of spinal cord injury by morphology, imaging andhistological detection means.ResultsThe enzyme digestion method could successfully separate, purify and culture human umbilicalcord mesenchymal stem cells. Detection and analysis with flow cytometry showed the thirdgeneration cells strongly express mesenchymal stem cell markers and(CD73, CD90, CD105) andadhesion molecular markers (CD54, CD44, CD29) with no expression of hematopoietic stemcell marker (CD45, CD40, CD34) and a sign of human leukocyte antigen (HLA-DR). Theenzyme digestion successfully separated, purified, and cultured rat Schwann cells identified bymorphology and optical GFAP immunocytochemistry. The built materials-cell complexes,through animal experiment,with imaging, histological and morphological detection showed goodeffect on spinal cord injury.ConclusionsThe successful construction of a method that can stabilize the separation, cultivation, proliferation of human umbilical cord mesenchymal stem cells and rat Schwann cells. The cellcomposite material constructed by the cultured cells of high purity, fast proliferation and stablecharacter and the chitosan-NT3, through the appropriate animal experiments, was confirmed tohave good effect on repair of rat spinal cord injury. |
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