A Atudy On Transplantation Of Human Umbilical Cord Derived-Mesenchymal Stem By Different Ways For The Treatment Of Spinal Cord Injury In Rats

Objective1.To establish the methods to isolate and proliferate human umbilical cord derivedmesenchymal stem cells (HUC-MSCs).2. To explore the curative effect of HUC-MSCsdirect local transplantation for repairing SCI in rats.3. To explore the curative effect ofHUC-MSCs intravenous transplantation for repairing SCI in rats and to observe themigration of HUC-MSCs into the injured parenchyma of spinal cord.4. To compare therepairing capability of HUC-MSCs transplantation via vein and local site injection forSpinal cord injure.5. To explore the possible mechanisms to repair spinal cord injury andlook for a appropriate transplant method for treatment of acute spinal cord injury inclinical.Materials and methods1. Isolation, culture, and identification of HUC-MSCs.HUC-MSCs were collected from full-term natural delivery, then Isolated, cultured,and identified.When the cells were passed the5th generation,we used Brdu to dark it. Afterthe growth of three days, digestived cells into the suspension readying to transplant.2. Preparation and transplantation in the treatment group of Spinal cord hittinginjury model.80wistar rats of clean grade, male or female, weighing200-220g, first the model ofspinal cord injury in rats were randomly divided into5groups: blank control group (groupA,10),DMEM local transplantation group (group B,15), DMEM transplantation groupby vein (group C,15),cells local transplantation group (group D,20),cells transplantationgroup by vein (group E,20). Group A only received simple injury,no transplantation;GroupB and C rats were transplanted in DMEM with the same amount volume of cells by local or vein;Group D and E rats were transplanted in suspension of HUC-MSCs by local andvein.3. The evaluation of recovery of spinal cord injury in rats.Observed and scored motor function of animal models in1days,1week,2weeks,3weeks,4weeks,6weeks,8weeks,10weeks and12weeks by BBB score. The formation ofhollow and the regeneration of axons through HE staining and silver staining, the survivaland distribution of labeled cells by immunohistochemical staining at random aftertransplantation. Imigration, differentiation and survive of HUC-MSCs in injury site wereobserved by immunohistochemistry. The area of glial scar in the injury site was calculatedby immunostaining against GAFP.ResultsBBB score of group D(7.20士0.70) and E(7.12士0.81) was more higher than groupA、B、C3weeks after injury, the statistical difference was significant (p <0.05). Thestatistical difference was not significant (p>0.05) between group D and E, while thestatistical difference was not significant (p>0.05) between group A and B and C. Thestatistical difference was not significant (p>0.05) between group A、B、C、D and E3d、1、2weeks after injury. The statistical difference was significant (p <0.05) between groupD、E and group A、B、C4、6、8、10、12weeks after injury. The statistical differencewas not significant (p>0.05) between group D and E, while the statistical difference wasnot significant (p>0.05) between group A、B and C.Observed by HE staining and silver staining, Group D and E had more regeneratingaxons, and part of nerve fibers passed through the injury zone, the difference ofregenerating axons in group D and E not significant. Group A、B、C had only few scatteredaxonal regeneration, nerve fibers of discontinuity passed through the injury zone.Immunohistochemical staining showed HUC-MSCs did not differentiate into neuronalcells, oligodendrocytes and astrocytes. The rats in Group E might see HUC-MSCs3weeksafter injury, and the rats in Group E might see a small amount of HUC-MSCs12weeksafter injury.Immunohistochemical staining showed lots of astrocytes can get through thedamage zone in Group D and Group E，tending to be normal, regularly arranged, scar areawas reduced compared with group A、B and C.Conclusion1. Can be isolate, culture and identificate HUC-MSCs in vitro. 2. HUC-MSCs local direct transplantation can restore neural function of rats withspinal cord injure.3. HUC-MSCs can migrate into the injure parenchyma of spinal cord and repair spinalcord injury via intrvenous transplantation.4. The therapeutic effect of HUC-MSCs intravenous transplantation for treatment SCIis similar to that by local direct transplantation.