Establishment Of Rat Amniotic Epithelial Cells Modified By BFGF And Effects Of The Cells On Optic Nerve Regeneration In Rat

In this study we established lentiviral vector with human bFGF gene,and successfully harvested highly infective viral supernatant after lentivirus packaging processes. Rat amniotic epithelial cells (AECs) were cultured in vitro and were identified with RT-PCR and immunocytochemistry.Then the AECs were infected with viral supernatant and were selected to obtain cells that express bFGF gene stably. RT-PCR,immunocytochemistry and ELISA methods were employed to identify expression and secretion of bFGF by constructed AECs. Furthermore,growth activity and neurotrophic activity of bFGF gene modified AECs was investigated. Then we implanted bFGF gene modified AECs and untransfected AECs into rat optic nerve transected by sucking. Hoechst33342 labeling, BDA labeling, Nissl staining and HE staining was employed to preliminarily observe the survival and migrative ablility of implanted cells,as well as the changes might happen on RGCs.Furthermore,the regeneration of injured axons was assessed by immunohistochemistry staining of GAP-43.Results implicated that bFGF gene modified AECs have much stronger growth activity and neurotrophic activity.Implanted bFGF gene modified AECs could survive massively within the injuried optic nerve and migrate into both directions to a relatively far distance,as well, the RGCs protective effect and a significant increase of cells within the distant part of the injuried optic nerve and better regeneration of the axon was observed. bFGF gene modified AECs could promote some of the axons regenerating within the injury area,and extending into the distal part about 0.3 mm,implicating that the cells could help to improve the microenvironment of the optic nerve.