关键词: 逆转录病毒载体;包装细胞;集落形成单位

    摘 要:目的 探讨产生重组逆转录病毒包装细胞系建立及其病毒滴度(以cfu表示)的影响因素. 方法 用逆转录病毒载体pSLXCMV，以磷酸钙共沉淀法转染PA317包装细胞，挑选抗性集落(PA317/pSLXCMV)扩大培养后，进行PA317/pSLXCMV细胞接种密度、培养温度(37℃，32℃)、纯化方法等对其培养上清中重组病毒cfu影响的比较性研究. 结果 包装细胞的密度是影响cfu滴度的关键因素;降低培养温度需延长培养时间方可提高cfu滴度;对比离心纯化与过滤纯化，并未发现两者之间的差异. 结论 该实验结果为应用逆转录病毒载体进行的ex vivo基因治疗实验研究提供重要参考指标.

    Optimized methods for preparation of viron produc┐ing package cell line

    ZHANG Hui-Zhong FAN Qing-Yu YANG An-Gang

    Department of Orthopaedics，Tangdu Hospital，Fourth Military Medical University，Xi'an710038，China，

    Department of Molecular Biology，Faculty of Preclinical Medicine，Fourth Military Medical Uni-versity

    Keywords:retroviral vector;packaging cell;colony-forming units

    Abstract:AIM To optimize the methods of manufacturing retrovirus containing supernatant of PA317packaging cell line for the use in the retroviral-mediated gene transfer.METHODS Studies were conducted using pSLXCMV retro-viral vector.The colony forming unit(cfu)influence factors including packaging cell density ......