Human Umbilical Cord Mesenchymal Stem Cells To Treatment Of Rat Spinal Cord Injury And Their Impact To Endogenous Stem Cells

BackgroundSpinal cord injury is a commen one of the central nervous system diseases,which is divided into two major categories:traumatic and non-traumatic, traumatic spinal cord injury in the majority,such as falling from height, transportthe accident caused spinal fractures often lead to serious damage to the spinal cord. Currently, the treatment of spinal cord injury is a worldwide medical problem, which is very complex pathological changes that occur after spinal cord injury, and neuronal damage are irreversible, and weaker in the regenerative capacity of neurons.Therefore, the treatment of spinal cord injury are facing many difficulties. Currently,the traditional treatment methods can not effectively solve the regeneration in the neuronal damage.In recent years, stem cells transplantation to repair sci have been becoming a hotspot. Many experiments with stem cells transplantation can effectively improve nerve function after spinal cord injury. Some country even made some clinical trial experiments by using stem cell transplantation therapy,which showed a ideal effect. hUCMSCs attracted much attention, its advantages compared with other mesenchymal stem cells is less ethical restrictions, low immunogenicity, easily isolated and cultured, and the study confirmed hUCMSCs with appropriate conditions can be differentiated into neuron-like cells. Therefore, hUCMSCs as a new seed cells for the treatment of nervous system injury. Transplantation of exogenous stem cells can affect endogenous stem cells in the central nervous system and the experiment is to study the neurological function recovery and the change of endogenous stem cells after spinal cord injury.ObjectiveTo observe the function recovery case of SD rats with spinal cord injury after accepting human umbilical cord mesenchymal stem cell transplantation, and what affect made to the endogenous stem cells.Materials and MethodsForty-six male SD rats (8weeks of age) provided by the Experimental Animal Center of Henan Province. All experimental course were performed according to guide for the care and use of laboratory animals(The National Ministry of Science and Technology,2006).All rats live under conditions of controlled temperature (23±2)℃and humidity (40%～60%).Isolation and culture of cells:In a sterile environment,get human umbilical cord from the post-partum period, cleaning, separation, cut it into pieces, and finally added to containing10%FBS (fetal bovine serum, FBS) and20g/L of alkaline into DMEM/F12, which also contain bFGF and were cultured and passaged:To observe the state of cells growth by the microscopy,when passaged to the the P4generation.SCI model making and grouping:Made the spinal cord injury model using modified Allen’s way. The experimental animals were divided into damage control group (PBS group)、normal group and transplantation group (hUCMSCs group), which hUCMSCs1group:efficacy group; hUCMSCs2:assessment group. All experimental rats were injected with BrdU (50mg/kg), once a day, a total of seven days for the newly generated cells labeled after transplantation. Transplanted rats with spinal cord injury48h, local injection around the spinal cord injury District, the control group in the same manner as the injection of the same amount of PBS solution.To evaluate the neurological function recovery of all rats by the BBB score; Observed the repair case of spinal cord injury by histological sections, and changes of endogenous stem cells by immunofluorescence labelin ResultsTissue cultured for7days, many hUCMSCs block out the tissue,and there were more cells adherent growth, long spindle shape,distributed evenly.About12days,about80%cells confluence, which can be passaged. One or two hours after the passage, cells began to attach to the cell wall, and stretch into polygons, gradually turned into a long spindle, evenly distributed in the sidewall. The cell proliferation latency shorter than the original, the proliferation rate of the cell passaging substituting fast.Damage control group compared to the transplantation group, in the7days after SCI and the49d, hUCMSCs group rated significantly higher than the PBS group (P <0.05); SCI after the49d, hUCMSCs group and PBS group scores were significantly higher than SCI7d (P<0.05). Histological staining can be observed in the damage zone of the hUCMSCs group voids smaller voids injury control group significantly larger, glial scar. Changes in endogenous cells:mesenchymal stem cell transplantation14d, the gray matter of the spinal cord and chamber tube membrane area of BrdU-positive cells were significantly higher than the PBS group (P<0.05). Immunofluorescence:7days after transplantation in spinal cord injury District around to observe to hUCMSCs groups the Nestin fluorescent activity stronger than the PBS group, and GFAP fluorescence activity is weaker compared to the PBS groupConclusionhUCMSCs transplantation promote functional recovery after spinal cord injury. Spinal cord injury after transplantation of hUCMSCs promote proliferation of endogenous stem cells.