Dissertation
Dissertation > Medicine, health > Basic Medical > Human morphology > Human histology

Infection and Latency of HSV-1 on Rat Bone Marrow Mesenchymal Stem Cells Vitro

Author PanLi
Tutor LiXiaoMian
School Tianjin Medical University
Course Pathogen Biology
Keywords Herpes simplex virus type 1 Latent infection Bone marrow mesenchymal stem cells Rats Tissue Engineering Organ transplant
CLC R329
Type Master's thesis
Year 2008
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Purpose: in vitro cultured rat bone marrow mesenchymal stem cells (BMSCs) establishment of bone marrow mesenchymal stem cells in vitro separation, culture conditions, explore bone marrow charge quality stem cells of the part of the biological characteristics, for further in-depth study bone marrow mesenchymal stem cells induced differentiation and to lay the foundation. Observation of herpes simplex virus type 1 infection of rat bone marrow mesenchymal stem cells and on the formation of latent infection, reveal the herpes simplex virus type 1 herpes simplex virus type 1 (HSV-1) infection of rat bone marrow mesenchymal stem cells of rat bone marrow mesenchymal stem cells. Method: under sterile conditions, stripped rat femur, tibia, since the rat femur, tibia bone marrow fluid lymphocyte separation medium separation of mesenchymal stem cells in the bone marrow, respectively, using conventional culture medium and culture medium containing mineralized solution two sets of cell culture, enzyme kinetics method, alkaline phosphatase (ALP) content detection and cell calcium nodules alizarin red staining, observation of mesenchymal stem cells in the bone marrow of the 3rd generation of bone marrow-derived mesenchymal stem cells in vitro long-term culture. calcium nodules formation of the situation, in order to do bone marrow mesenchymal stem cells identified; HSV-1 infection of bone marrow mesenchymal stem cells, the observed virus induced cell lesions (CPE); establishment of bone marrow mesenchymal stem cells of the HSV-1 latent infection model. Extraction of total DNA, PCR Amplification of bone marrow mesenchymal stem cells within the HSV-1 specific fragments, detection of HSV-1 infection of bone marrow mesenchymal stem cells and latent infection. Results: The two groups of bone marrow mesenchymal stem cells the culture liquid content of the ALP rising after two independent samples t test, the mineralization fluid induced group 3,6,9 ALP levels than normal control group were significantly increased (P lt; 0.05). Bone marrow mesenchymal stem cells after 14 days after the induction of the mineralized solution, cells exhibit characteristics of osteoblasts, the formation of calcium nodules in vitro long-term subculture. HSV-1 infection of rat bone marrow mesenchymal stem cells, a typical cell lesions (CPE) PCR method confirmed the bone marrow mesenchymal stem cells exist in HSV-1 specific fragments. HSV-1 latent infection of bone marrow mesenchymal stem cells did not appear significant cell lesions (CPE), bone marrow mesenchymal stem cells spread to the 7th generation, can still be measured to of HSV-1 gene fragment showed that bone marrow mesenchymal stem cells There is a possible form of HSV-1 latent infection. Conclusion: rat bone marrow mesenchymal stem cells in vitro culture conditions, a strong proliferative capacity, differentiation to osteoblasts direction, can be used as seed cells for tissue engineering. That bone marrow-derived mesenchymal stem cells are easy to get, easy to train, low immunogenicity, easily into and expression of exogenous gene characteristics, has become an effective carrier has potential practical value of the cell therapy and gene therapy clinical disease. HSV-1 in vitro infection of rat bone marrow mesenchymal stem cells have a latent infection trends. In this study, the clinical application of bone marrow mesenchymal stem cells for cell transplantation, gene therapy and organ transplantation, clinical medicine and improve the quality of life and survival of transplant patients, can provide a certain basis.

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