摘要
①目的 比较逆转录病毒载体 4种转染方法对小鼠脾脏T淋巴细胞的转染率。②方法 应用衣霉素预处理包装细胞提高逆转录病毒滴度 ,运用标准法、离心法、共培养法和脂质体 病毒混合法转染T淋巴细胞。③结果 共培养法转染率为 32 .33%± 2 .1 2 % ,离心法为 1 8.72 %± 2 .97% ,脂质体 病毒混合法为 9.4 3%±1 .81 % ,标准法转染率为 6 .6 4 %± 1 .37%。共培养法转染率高于离心法 (F =95 .2 4 ,q =35 .0 6 ,P <0 .0 1 ) ;共培养法和离心法转染率均高于标准法 (q =1 8.79、1 7.5 9,P <0 .0 1 ) ;脂质体 病毒混合法和标准法的转染率无显著差异(q =3.1 1 ,P >0 .0 5 )。共培养法转染率最高 ,但易于混入包装细胞。
Objective To compare the transducting efficiencies of four transduction methods through transfecting murine splenic T lymphocytes with retroviral vectors. Methods Packaging cells were pre-treated by tunicamycin to improve retroviral vector titer. Splenic T lymphocytes were transfected with four kinds of transduction methods, i.e. standard, centrifugation,cocultivation and virosomes transduction. Results Transducting efficiency was as follows: cocultivation, 32.33%±2.12%; centrifugation,18.72%±2.97%; virosomes, 9.43%±1.81%; and standard transduction 6.64%±1.37%. Transducting efficiency of cocultivation was higher than that of centrifugation ( F=95.24,q=35.06,P <0.01), and those of both cocultivation and centrifugation were higher than that of standard transduction( q=18.79,17.59,P <0.01). There was no significant difference between virosomes and standard transduction ( q=3.11,P >0.05). Despite the highest transducting efficiencies, cocultivation transduction was prone to mix T lymphocytes with packaging cells. Conclusion Centrifugation transduction was a preferred transducting method.
出处
《青岛大学医学院学报》
CAS
2004年第2期131-133,136,共4页
Acta Academiae Medicinae Qingdao Universitatis
