Influence Of The Latency Of Herpes Simplex Virus Type1on Bone Marrow Mesenchymal Stem Cells Transplantation

Objective:Separated and cultured of rat bone marrow-derived mesenchymal stem cells (BMSCs) in vitro, to study its biological characteristics and culture methods, in order to lay the foundation for further experimental study. Infected mice by herpes simplex virus type1(HSV-1), to study the change of its behaviory, whether HSV-1could cause latent infection on the host bone marrow and HSV-1is persistence with the differentiation of BMSCs; To study whether HSV-1could persistence consistently after the transplantation of BMSCs with the latency of HSV-1by implanting BMSCs that carried HSV-1. It is warning for the bone marrow or stem cell transplantation, clinical application and so on.Method:We isolated and cultured BMSCs by wall adhesion culture procedure and density gradient centrifugation, and compared the cell growth rate, adhesion rate, multi-directional differentiation functions and other cell characteristics. Mice of the experimental group and control group were sacrificed respectively in the5th day, the15th day and the30th day after tail vein injection of HSV-1, and the primary BMSCs were cultured and identified. The specific gene fragments of HSV-1were examined by Polymerase chain reaction in bone marrow and BMSCs. Rats of the experimental group and control group were sacrificed respectively in the2nd day, the4th day and the6th day after implanting BMSCs which carried HSV-1. The specific gene fragments of HSV-1were examined by Polymerase chain reaction in BMSCs and bone marrow.Results:The proliferation and activity of BMSCs prepared by adherent selection are much higher than those prepared by density gradient centrifugation. But there is no difference in other cell characteristics, such as multi-directional differentiation function and cell morphous. The cells obtained from BMSCs were identified to be osteoblasts by alkaline phosphatase stain and alizarin bordeaux stain of calcified nodule; The typical morphology of dendritic cells which also obtained from BMSCs were cultured. The specific gene fragments of the experimental group and its corresponding first and second generation BMSCs were amplified correctly by Polymerase chain reaction. In BMSCs transplantation experiments, the specific gene fragments of the BMSCs and bone marrow in each experimental group were amplified correctly by Polymerase chain reaction.Conclusion:Among the methods of BMSCs preparation, pastes the wall screening law is the most economical and practical method. Pure BMSCs could be got by density gradient centrifugation, which is also a commonly used method. This experiment confirmed that bone marrow is one of the organs that could be infected by HSV-1which could consistently exist with cell proliferation, and HSV-1could be reactivated after the transplantation of BMSCs with the latency of HSV-1. There were potential risk factors in bone marrow transplantation and clinical application of BMSCs currently, for exmple, donors and patients could carry HSV-1.This proposed warning for bone marrow transplants and associated clinical application, excluding HSV-1infection is needed.