体外培养表达人β-NGF基因的猫角膜内皮细胞

Research on cat corneal endothelium cells expressed human β-nerve growth factor gene cultured in vitro

目的:体外培养表达人β-神经生长因子基因(β-nerve growth factor,β-NGF)的工程化猫角膜内皮细胞,为进一步猫角膜内皮细胞的移植做准备。方法:将人β-NGF重组真核表达载体pcDNA4-β-NGF,通过EffecteneTM脂质体介导转染到体外培养的猫角膜内皮细胞中,选择性培养基筛选转基因后细胞,人NGF单克隆抗体行细胞免疫组织化学染色(immunohistochemistry)。结果:选择性培养基培养2wk后筛选出独立细胞克隆,该细胞高表达人β-NGF基因。结论:转基因方法可体外培养出表达人β-NGF基因的工程化猫角膜内皮细胞。

AIM： Cat corneal endothelium cells which expressed human β-nerve growth factor （β-NGF） gene were cultured in vitro for preparation of cat corneal endothelium cells transplantations. METHODS： Human β-NGF recombination eukaryotic expression vectors were transferred into the cat corneal endothelium cells by EffecteneTM liposomeo Positive expressed cell clones were sieved out by selecting culture medium, human NGF monoclonal antibody cell immunohistochemistry staining was used by human NGF monoclonal antibody. RESULTS： After cultured in selective culture medium for 2 weeks, solo cell clones were sieved out, and the cell highly express the human β-NGF gene. CONCLUSION ： Cat corneal endothelium cells expressed human β-NGF gene could be cultured in vitro by transgenic methods.