Human nerve growth factor beta-modified bone marrow mesenchymal stem cells from the rabbit mandible by transfection of lentiviral vectors / 中国组织工程研究

ABSTRACT

BACKGROUND:Central nerve damage and peripheral nerve injury are common clinical problems that have no ideal treatment. Nerve growth factor has an important role in neuronal repairing and growth. But its local injections may have shorts of inactivation and loss. OBJECTIVE:To construct human nerve growth factor beta recombinant plasmids, which are transfected into bone marrow mesenchymal stem cells from the rabbit mandible by lentiviral vectors, and to investigate the bioactivity of human nerve growth factor beta. METHODS:pDC316-hNGFβ-mCMV-EGFP plasmids were constructed via lentiviral vectors using Hind III+Not I digestion. Bone marrow mesenchymal stem cells from the rabbit mandible were isolated and cultured, and then transfected by recombinant plasmids. The expression of human nerve growth factor beta in transfected cells was detected by ELISA method. RESULTS AND CONCLUSION:pDC316-hNGFβ-mCMV-EGFP plasmids were proved to be constructed successful y by gene sequencing and enzyme identification. The transfected cells under a fluorescence microscope emitted green fluorescence, and the fluorescence intensity had no change with incubation time. The expression of human nerve growth factor beta was maintained at a level of 25μg/L at 7 days after celltransfection, and the bioacitivty was increased significnalty.