Study On Human Umbilical Cord Blood-derived Mesenchymal Stem Cells Can Be Induced To Neuron-like Cells And Transplantation

Objective: 1. To establish the method of in vitro culture and expansion of human umbilical blood-derived mesenchymal stem cells (MSCs) and to investigate their biological characteristics.2. To investigate the feasibility of using MSCs(Mesenchymal Stem Cells)derived from HUCB(Human umbilical Cord Blood) to regenerate neuron like cells and to explore the optimal condition of isolation, purification and amplification in vitro.3. To study whether intraspinally transplantation of using MSCs(Mesenchymal Stem Cells)derived from HUCB(Human umbilical Cord Blood) induction differentiation into regenerate neuron like cells ,and improve neurological functional recovery after spinal cord injury in rats. Methods: 1.HUCB were collected from full term deliveries scheduled for cesarean section, the cord blood mononuclear cell were isolated by lymphocyte separation medium, purified and culture with MesencultTM medium and acidic environment to produce adherent layer, the growth curves of MSCs were drawn.2. HUCB were collected from full term deliveries scheduled for cesarean section, all specimens were obtained sterilely with preservative –free heparin, the cord blood mononuclear cell were isolated by lymphocyte separation medium, purified and expanded with MesencultTM medium and acidic environment to produce adherent layer, after the passage 3 of the expanded of MSC were induced to differentiate to neuron like cells. Specific marks of neuron were detected by fluorescent Immunohistochemical stain method.3. Rats were randomly divided into two groups.Subjected to NYU spinal cord injury pattern.One group of rats was transplanted with MSCs(Mesenchymal Stem Cells)derived from HUCB(Human umbilical Cord Blood) induction differentiation into regenerate neuron like cells labeld by bromodeoxyuridine(BrdU).The other group was injected to PBS(control group).The neurological function was determined before and 24h,1,2,3,4 and 5,weeks after spinal cord injury and cell transplantation using the modified BBB score.The distribution and differentiation of transplanted cells in vivo in rat spinal cord were evaluated by histological and immnuhistochemical analysis. Results: 1.The HUCB-derived mononuclear cells, when seed in suitable medium, gave rise to adherent cells, which exhibited either an osteoclast-or mesenchymal-like phenotype, cells with the MSCs displayed a fibroblast-like morphology and expressed several MSCs-related antigens which was identical to human bone marrow-derived MSCs.2. The HUCB-derived mononuclear cells, when seed in specific medium, gave rise to adherent cells, which exhibited either an osteoclast-or mesenchymal-like phenotype. After passage 3, these cells can be purified and expanded. Immunohistochemistry staining showed that the induced MSCs expressed human glial fibrillary acidic protein(GFAP),neurofilament(NF) and neuron-specific enolase(NSE). Conclusion: 1.MSCs in HUCB can be cultured and expanded in vitro, and could be regarded as an alternative source of MSCs for experimental and clinical applications.2. In this study shown MSCs in HUCB can be cultured and expanded in vitro, and could MSCs(Mesenchymal Stem Cells)derived from HUCB(Human umbilical Cord Blood) induction differentiation into regenerate neuron like cells ,and improve neurological functional recovery after spinal cord injury in rats.be a source of stem cells which can be differentiated into neural like cells for experimental and clinical application.3. MSCs(Mesenchymal Stem Cells)derived from HUCB(Human umbilical Cord Blood) induction differentiation into regenerate neuron like cells ,and improve neurological functional recovery after spinal cord injury in rats.