重组人生长激素基因细胞移植

Implantation of transfected the human growth hormone gene fibroblasts into mice: implications for gene therapy

目的 通过重组人生长激素基因细胞移植为生长激素缺乏症 (GHD)基因治疗的生物学表达研究奠定基础。方法 构建 pLXSNhGH人生长激素 (hGH)逆转录病毒表达载体后 ,脂质体转染包装细胞系PA317,提取包装细胞上清的病毒后 ,将病毒感染原代小鼠胚胎成纤维细胞 ,将细胞移植于小鼠腹腔内 ,观察hGH的表达情况。结果 hGH体内持续表达达 2个月以上 ,但由于高滴度抗hGH抗体的影响 ,表达水平呈波动性。结论 重组人生长激素基因原代成纤维细胞移植体内的持续表达 。

Objective To lay the foundation of biological expression studies of growth hormone deficiency gene therapy with transplanting cell of recombinant human growth hormone gene in vivo. Methods After construction of a human growth hormone retrovirus expression vector pLXSNhGH, The recombinant vector was transfected into packaging cell lines PA317 via lipofection, G418 select resistant colony, the virus were collected from the virus containing supernatant of packaging cell lines, primary mouse embryo fibroblast cells were infected. The cells were transplanted at donor intraperitoneal sites and were detected in the donors circulation. Results Fluctuating levels of human growth hormone were detected for over 2 months due to interference by high titers of antibodies developed against the human growth hormone. Conclusions Primary fibroblasts tranfected by recombinant human growth hormone gene can produce novel gene products lasting for over 2 months, which lay the foundation of growth hormone deficiency gene therapy in vivo.