BFGF Gene Transfection And Expression Of Its MRNA And Protein In Schwann Cells

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 reconstruct a new plasmid of bFGF- pcDNA3.1 and then transfected it into the schwann cell (SC) by liposome so that to achieve the high and stable expression of bFGF .That both supply abundant bFGF and improve the local minimal environment.Our research contains 3 parts1. Obtain the highly purified schwann cell.2. reconstruct the plasmid bFGF- pcDNA3.13. Transfected the pcDNA3.1-bFGF into the schwann cell (SC) by liposome, then test the bFGF expression level in the schwann cell culture supernatant of experiment group, empty carrier group and control group respectively by ELISA . Exam the bFGF mRNA expression by RT -PCRResults1.Highly purified schwann cell can be obtained from mouse sciatic nerve by pre-degeneration, coenzyme digestion and G-418 bolting. 2.the eukaryotic expression system of destination gene and activity is reconstructed by bFGF and pcDNA3.1 that is of high security and replica.3. After bFGF- pcDNA3.1 has been transfected into schwann cell by liposome, the prompt, persistant, stable and high expression of bFGF can be obtainedConclusion :1. bFGF can be expressed and excreted into the supernatant after bFGF- pcDNA3.1 transfected into schwann cell by liposome.2. schwann cell modified by bFGF- pcDNA3.1 can express and excrete bFGF highly and stably ,but we also observe the reduction of bFGF expression when schwann cell is developed in vitro due to the limitation of culture environment.3. It is of unique advantages to connect schwann cell and bFGF- pcDNA3, which not only expresses bFGF enough and stably in trauma spine but supports and leads to the neuro-axon regeneration. Therefore, it offers the significant conditions for the damaged spinal neuron's rehabilitation and regeneration and shows the bright future for the clinical treatment of spinal injury.