摘要
应用逆转录病毒载梧介导基因转移法将ｎｅｏ＾ｒ基因导入人造血干祖细胞，并经体外液体长期培养。结果表明，产重祖病毒细胞ＰＡ３１７／Ｎ２的病毒滴度最高达６．５×１０＾５ＣＦＵ／ｍｌ，用ＰＣＲ法从体外培养２，４，６Ｗ及加入ｒＩＬ－１，ｒＩＬ－６的悬浮和贴壁的转染细胞中均可扩增出ｎｅｏ＾ｒ基因ｃＤＮＡ片段，非同位素化学发光法标记ｎｅｏ＾ｒ探针与之杂交显示阳性结果。
Retroviral vectors containing the bacterial neomycin resistant(neo￣r) gene were transfered into primary human hematopoietic progenitor cells, and were cultured into long-term repopulating marrow cells(LTC).The results showed that was obtained a high titer (6.5× 105 CFU/ml)helper-free,virus producing cells-pA317/N2.The neo￣r gene from both noadherent and adherent LTC cells which have been cultured from 2、4、6 w and added rIL-1 ,rIL6 was detected directly by PCR. The direct labelling-chemilumcium scent photographic was used to detect the neo￣r gene transcripted and expressed in LTC. These studies demonstrate that the neo￣r gene can be successfully transfected and expressed in human hematopoietic stem progenitor cells of LTC at high efficiency using the retrovirus vectors.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
1996年第6期335-338,共4页
Chinese Journal of Immunology
关键词
逆转录病毒载体
造血干祖细胞
基因转移
Retroviral vector Human hematopoietic progenitor cell Long-term marrow culture Gene transfer
