A Study Of Transplantation Of HUCBMSCs Result In Effect In The Spinal Cord Of Adult Rats After Spinal Cord Injury

Bankground and Objective:At present, mesenchyme stem cells (MSCs) in different tissue can be see as Adult stem cells (ASCs) from different tissues in nature. It can not only pack proliferate in-vitro-cultivation conditions, but also keep the state of extreme undifferentiation. MSCs have the multilineage differentiation potential , and ,by targeting inducing differentiation in virto, they can transform into osteoblast, chondroblast, sarcoblast, adipocyte, vascular endothelial cell,etc. In the suitable conditions, MSCs can convert neural cells too.Near decade, how get the great deal of MSCs to suffice the clinical demanding had become the focus of the study on stem cells. Anciently, many researchs use the bone marrow mesenchyme stem cells as the material and transform these cells into neural cells in virto or directly transplant them into the rats' brain, which have succeeded in resuming the function of the-stroke-rats. Bone marrow mesenchyme stem cells have some disadvantages such as: The source was limited largely and it's very possible that have infected virus.With the years going, the number of the cells and the ability of amplification and differentiation will decrease and degression. For these reasons , on the same time, For obtaining enough research cells , people start to research the feasibility and the method of separating and cultivating the multilineage differentiation potential MSCs from the human umbilical cord blood . certainly, human umbilical cord blood MSCs could be supposed to replace the bone marrow MSCs and make up it's defect too. From a lot of research ,people find that the human umbilical cord blood and the bone marrow both contain the MSCs, but it was more original and have better ability of multiplication and differentiation that the MSCs derive from the human umbilical cord blood than from the bone marrow. This made a new choice for the source of transplant.HUCBMSCs were separated, cultured, proliferated, and then implanted into the rats of Spinal Cord Injury. Comparing with other groups, the effect of Spinal Cord Injury cured by transplantation of HUCBMSCs was evaluated by behavioral score . This experiment may give experimental and academic theory base for the clinical use of HUCBMSCs as a new source of cells for cellular therapeutic for diseases in central nervous system.Object:To find the effect of Spinal Cord Injury cured by transplantation of HUCBMSCs and give experimental and academic theory base for the clinical use of HUCBMSCs.Methods:Mononuclear cells (MNCs) were prepared from heparinized human umbilical cord blood samples (donated from the full term volunteers) by centrifugation over Lymphoprep (1.077 g/ml) and cultured in Mesencult^TM , then got the MSCs through purification and amplification.Total 46 adult healthy Wistar rats were randomly allocated into 3 groups: A groups were SCI, B groups were SCI+NS and C groups were SCI+HUCBMSCs. Group A and B has 15 and C has 16 rats. All the rats models of SCI were established by applying a free-falling device hiting the rats' brain directly. Three groups all were evaluated by behavior in the injury's days ,7 days ,14days and 4weeks after transplantation. The evaluation standard was Basso-Beatie-Bresnehan grade. Inflammatory reaction was observed in HE slice in 1 week and 4 weeks after transplantation. GFAP, NSE were tested according to immunocytochemistry method in 4 weeks after transplantation. The experimental group and the control group would be contrasted.Result: The MNCs from the human umbilical cord blood which was implanted into special culture medium could produce adherence-cells and these cells mainly presented osteoclast-like cells and mesenchymal-like cells. Through three passages, purification and amplification would be reached. These cells mostly would present mesenchymal-like cells. HUCBMSCs transplantation could improve posterior limb motive function. It could be observed after two weeks and would become more obvious after four weeks on BBB grade. there was significant diference (P <0.05) between C groups and B groups, A groups in the scores of BBB grade. Immunological rejection wasn't visible after HUCBMSCs transplantation. There were great of HUCBMSCs survival in 4w after transplantation and it could be observed that some new glial cell formed.Conclusions:1. HUCBMSCs can be well cultivated in Mesencult^TM.2. HUCBMSCs transplantation can achieve satisfying effect of treating SCI rats.3. HUCBMSCs could survival and form some new glial cell after transplantation.