BFGF Gene Modified MSCs And Of Its MRNA And Protein Express In SCI Animal Model

Spinal cord injuries often characterized by immediateand irreversible loss of sensor and motors below the level of injury,and exacta physical, emotional and economic toll both on those who are affected andon society at large.The capacity of certain damaged axons to regeneratationand elongation in the CNS and PNS are more dependent on the enviroment in which these axons are located than intrinsic properties of neurons.Celltransplantation to repair spinal cord injury is a vibrant area of research with the goal of reducing functional defrcit,which can facilitates growth of axons by serving as a celuar bridge and providing chemical and mechanical cues for injured neural processes,provides new neurons which in turn provide new targets and sources of innervation and repair damaged neural circuits, and secretes an array of factors,such as neurotrophic factors,that might aid in the repair process.bFGF (FGF-2) (basic fibroblast growth factor) is one member of FGF family which related to the angiogeneration , fetus growing and the neuro-generation. bFGF generally exists throughout the body especially in nerves. Nowadays, lots of the investigations have shown the significance of bFGF at the spinal regeneration and rehabilitation after injury. However ,after acute spinal injury, bFGFR expresses earlier than bFGF which diminishes its protection for the ischemic neuron. Therefore, during spinal injury treatment, the abundant bFGF supply before bFGFR expression reaches its climax is of the significant importance. But, unfortunately, neither local nor general application of the external bFGF can reach the ideal result. Based on the fact above mentioned ,our research is trying to make some improvements. We Transfect the pcDNA3.1-bFGF into the bone marrow stromal cell (MSCs) by liposome, so that to achieve the high and stable expression of bFGF in body .That both supply abundant bFGF and improve the local minimal environment.The article establish the foundation for the post research. Our research contains 3 parts1.Obtain the highly purified bone marrow stromal cell(MSCs) by screeing-adherence methord.2.Transfect the pcDNA3.1-bFGF into the bone marrow stromal cell (MSCs) by liposome, then test the bFGF expression level in the bone marrow stromal cell .3. Transplant bone marrow stromal cell modified by bFGF- pcDNA3.1 to SCI animal model.Test bFGF protein expression in gene group,empty carrier group,MSCs group and DMEM group respectively by immunohistochemistry; Exam the bFGF mRNA of each group expression by RT -PCR .Result of the reseach1.Highly purified bone marrow stromal cell can be obtained from mouse bone by screeing-adherence methord.2.After bFGF- pcDNA3.1 has been transfected into bone marrow stromal cell by liposome, the effective expression of bFGF can be obtained3. After gene modified MSCs has been transplanted into SCI animal model, the prompt, persistant and stable expression of bFGF can be obtained ,that elevate the level of bFGF expression.Conclusion :(1)we can obtain highly purified bone marrow stromal cell by screeing-adherence methord. (2)The effective expression of bFGF can be obtained ,after bFGF- pcDNA3.1 has been transfected into bone marrow stromal cell by liposome. (3)It is prevalent to transplant MSCs together with bFGF gene,which supply spinal cord injuried with stable and high expression of bFGF protein.