[关键词]
[摘要]
目的：构建过表达睫状神经营养因子(CNTF)的腺病毒，利用腺病毒载体转染骨髓间充质干细胞，使其在体外持续高效表达CNTF，为视网膜病变的体外研究提供新途径。
方法：构建空载GFP-腺病毒及CNTF-腺病毒，取第3代BMSCs用于转染实验，分为3组：空白对照组(未转染腺病毒组)、阴性对照组(GFP-腺病毒转染组)和实验组(CNTF-腺病毒转染组)。ELISA测转染后1、2、3d各组上清液的CNTF蛋白分泌量。
结果：成功构建空载GFP-腺病毒及CNTF-腺病毒， 实验组的CNTF表达量明显高于空白对照组和阴性对照组，差异具有统计学意义(P<0.05)。
结论：利用腺病毒载体可使BMSCs在体外持续高效表达CNTF。
[Key word]
[Abstract]
AIM: To establish the model of bone marrow mesenchymal stem cells(BMSCs)over-expressing ciliary neurotrophic factor(CNTF)by applying adenovirus. It may provide novel strategies for
in-vitro investigations of retinopathies.
METHODS: GFP-adenovirus and CNTF-adenovirus were synthesized then used to transfect BMSCs of passage 3. Blank control group(without adenovirus transfected group), negative control group(GFP-adenovirus transfected group), and experimental group(CNTF-adenovirus transfected group)were included in this study. On the 1, 2, 3d post-transfected, ELISA assay was applied to examine CNTF protein-secretion in the supernate.
RESULTS: GFP-adenovirus and CNTF-adenovirus models were successfully established. The CNTF protein levels in the supernate were higher in experimental group than those in the blank control group and negative control group(P<0.05).
CONCLUSION: CNTF-modified BMSCs by adenovirus could efficiently secrete CNTF protein in-vitro.
[中图分类号]
[基金项目]
国家自然科学基金资助项目(No.81770948); 福建省科技创新领军人才基金资助项目(No.2016B011)
