论文部分内容阅读
目的 探讨小鼠胚胎干细胞 (TC 1)转基因治疗甲状旁腺功能低下症 (HPT)。方法包装出重组人甲状旁腺素 (PTH )基因的小鼠干细胞病毒 (MSCV) ,以lml重组病毒液加入 poly brene(终浓度 8mg/L)感染TC 1细胞 ,检测基因转导效率 ,PTH分泌情况 ;以及每 1× 10 5个 /mlTC 1细胞注入模型鼠体内各组小鼠血PTH和血钙变化情况。结果 获得滴度为 2× 10 7集落形成单位 (CFU) /ml的重组逆转录病毒 ,其感染TC 1的效率为 70 % ,每 10 6 未分化TC 1每 48h分泌PTH10ng。重组有PTH基因的TC 1细胞注入模型鼠体内后 ,在观察期间实验组动物血PTH和血钙均保持在接近正常值范围内。结论 MSCV介导外源PTH基因可高效转导TC 1并持续分泌PTH ;内环境并不是决定TC 1分化的唯一因素。经基因转导的TC 1可较好的改善模型鼠的症状 ,是未来细胞移植的一种潜在来源。
Objective To investigate the effect of transfection of mouse embryonic stem cell (TC 1) on hypoparathyroidism (HPT). Methods Recombinant human parathyroid hormone (PTH) gene murine stem cell virus (MSCV) was packaged and inoculated with poly brene (final concentration 8 mg / L) in 1 ml of recombinant virus solution to infect TC1 cells. The gene transduction efficiency, PTH secretion And the change of blood PTH and blood calcium in each group of mice injected with 1 × 10 5 / ml TC 1 cells. As a result, a recombinant retrovirus having a titer of 2 × 10 7 colony forming units (CFU) / ml was obtained, the infection rate of TC 1 was 70%, and PTH was secreted every 10 6 undifferentiated TC 1. After the PTH-transfected TC1 cells were injected into the model mice, the levels of PTH and serum calcium of the experimental animals remained close to their normal values during the observation period. Conclusion The MSCV-mediated exogenous PTH gene can efficiently transduce TC 1 and continue to secrete PTH. The internal environment is not the only factor that determines the differentiation of TC 1. Gene-transduced TC 1 can better improve the symptoms of model mice and is a potential source of future cell transplantation.