重组人生长激素基因细胞移植

The construction and in vitro expression of the human growth hormone gene-containing recombination vector

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。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,G418筛选抗性克隆 ,扩增单个克隆后提取包装细胞上清的病毒 ,将病毒感染原代小鼠胚胎成纤维细胞 ,将此细胞移植于小鼠腹腔内 ,观察hGH的表达情况。结果 :hGH体内持续表达长达两个月以上 ,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 ,为细胞移植系统引入GHD基因治疗的研究奠定了基础。

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 was detected in the donor 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 humangrowth hormone.Conclusions:Primary fibroblasts of recombinant human growth hormone gene were delivered novel gene products lasting for over 2 months,which lay the foundation of growth hormone deficiency gene therapy in vivo.\;