体内转染人突变DHFR原小鼠骨髓对致死量照射小鼠造血重建和 …

目的 将转染了人突变ＤＨＦＲ基因的小鼠骨髓移植给经致死剂量照射的小鼠，观察棋 对受者造血功能的重建和保护作用。方法 分离转染人突变ＤＨＦＲ基因小鼠骨髓有核细胞，移植给经致死剂量照射的同系小鼠，以甲氨蝶呤（ＭＴＸ）筛选．观察受体小鼠血象、生存率和ＣＦＵ－ＧＭ的改变，并用ＰＣＲ和Ｓｏｕｔｈｅｒｎ印迹杂交分析外源基因在小鼠染色体ＤＮＡ中的整合与表达情况。对照组以正常同系小鼠为供体。结果 在大剂量ＭＴＸ筛

Reconstitution and protection of murine hematopoiesis by second transplantation with bone marrow cells containing mutant human DHFR gene from primary generation murine

Objective To observe the effect of donor murine bone marrow, which was transfected in vivo with mutant hDHFR gene, on the reconstitution and protection of hematopoiesis of lethally irradiated recipient mice. Methods Mice bone marrow transduced in vivo with mutant hDHFR gene were used for transplantation. After whole bodily irradiated (8.59.0Gy) and then transplanted with the transduced marrow, the recipients were treated with methotrexate (4mg/kg twice a week for the first week; and then 10mg/kg twice a week). The survival rate and blood picture of the recipients as well as the integration and expression of hDHFR gene were studied by using PCR and Southern blotting. Results The hematopoiesis of the recipient mice could be reconstituted and protected from lethal toxicity of high dose methotrexate. It was confirmed that hDHFR gene was integrated into hematopoietic cell as well as some tissues genomic DNA and expressed successfully in the recipients. ConclusionDonor mice bone marrow transfected with mutant hDHFR gene can effectively reconstitute and protect hematopoiesis in lethally irradiated recipient mice.