基因修饰骨髓细胞移植后重建造血的实验观察

Experimental studies on hematopoiesis reconstitution by using of transplantation of gene modified bone marrow cells.

目的 探索利用基因标记技术观察造血重建的生物学特征。方法 利用脂质体介导将新霉素抵抗（ＮｅｏＲ）基因转导进入小鼠骨髓细胞。然后移植给受致死量照射的同种小鼠。观察受体小鼠造血的重建， 并对造血重建后受体鼠的脾及骨髓细胞进行标志基因的检测。结果 移植小鼠健康存活， 并形成脾结节。而未经输注的对照鼠则很快死于骨髓衰竭。同时， 移植鼠的部分脾及骨髓细胞于Ｇ４１８ 体系中能够存活， 经聚合酶链反应（ＰＣＲ） 检测含有ＮｅｏＲ 基因片段。结论 基因修饰骨髓细胞能够成功地重建造血， 并在一定程度上稳定地表达外源基因。

Objective To explore the biological features of hematopoiesis reconstitution by using genetic marking.Methods Neo R gene was transduced into bone marrow (BM) cells of mouse mediated by liposome. Then these cells were transplanted into lethally irradiated recipients. The hematopoiesis reconstitution was observed and the marker gene in spleen and BM cells of recipients after hematopoiesis reconstitution was examined.Results The transplanted recipients remained alive and healthy. But the irradiated mice with no transplantation died from BM aplasia soon. Meanwhile, the cells from spleen and BM of transplanted mice could be alive in G418 system, and contained the DNA fragments of Neo R gene by PCR. Conclusion Gene modified BM cells could be used to reconstitute hematopoiesis successfully and express the foreign gene to some extent stably.