pIRES2-EGFP-IGF-1真核表达载体转染骨髓基质细胞的动物实验

目的构建表达胰岛素样生长因子1（IGF-1）的骨髓基质细胞（MSC）的动物模型，为进一步将其用于修复牙周组织缺损奠定基础。方法将真核表达载体pIRES2-EGFP—IGF-1转染MSC，获得阳性细胞克隆，免疫组化、Western blot鉴定IGF-1蛋白表达水平，ELISA方法检测转染细胞上清液中IGF-1蛋白含量；用转染细胞上清液培养牙周膜细胞（PDLC），甲基噻唑基四唑（MTT）法检测对细胞增殖的影响。结果免疫组化和Westernblot结果证实，转染细胞内有IGF-1蛋白的高表达。ELISA方法检测显示，IGF-1能分泌至细胞外，且能明显促进PDLC的增殖。结论IGF-1基因可成功

Experimental study on dog''''s bone marrow stem cells transfected by pIRES2-EGFP-IGF-1 gene

OBJECTIVE: To establish the bone marrow stem cells (MSC) model which could highly express the insulin-like growth factor 1 (IGF-1) transfected by dog's IGF-1 gene. METHODS: pIRES2-EGFP-IGF-1 was transfected into MSC by lipofectamine. Positive clones were selected with G418. The expression of IGF-1 protein in the MSC was determined by immunohistochemistry and Western blot analysis. The IGF-1 in the supernatant of the transfected MSC was detected by sandwich-in ELISA. The periodontal ligament cells (PDLC) were cultured in the supernatant of the transfected MSC. The changes of PDLC' proliferation were observed by MTT. RESULTS: IGF-1-transfected MSC could apparently express IGF-1. The IGF-1 protein in the supernatant of the transfected MSC was confirmed by sandwich-in ELISA. IGF-1 could promote the PDLC' proliferation. CONCLUSIONS: The MSC transfected by dog's IGF-1 gene can highly express IGF-1, which may lay the foundation for further study on periodontal regeneration.