Sry监测体外扩增造血细胞植入效果的实验研究

目的：探讨Y染色体性别决定基因(Sry)作为评价和追踪造血细胞移植后植入状态检测指标的可行性。方法：根据Sry DNA序列设计引物及制备探针，然后进行PCR检测、斑点杂交和原位杂交．动态追踪经不同条件下体外扩增的纯系雄性小鼠BMMNC回输至雌性小鼠后的植入状态。结果：PCR结果显示在各移植组小鼠的骨髓有核细胞、脾细胞及外周血白细胞中均有Y染色体特异性序列的存在；斑点杂交结果显示在各回输组间并无明显差别，但用脾脏组织作原位杂交的结果则发现基质细胞支持扩增回输组的阳性颗粒数目与输注新鲜细胞组相似，但略少于雄性小鼠；而输注细胞因子扩增细胞实验组小鼠则明显少于雄性动物的阳性对照标本和基质细胞支持扩增回输组。结论：(1)经重复检测表明上述方法的重复性好，结果可信．可作为检测性别不同时造血干细胞移植效果的一种检测手段；(2)证实基质细胞支持下的体外扩增的造血细胞具有较好的植入能力。

The study on evaluating Sry as a engrafting track of the bone marrow cells expanded under the different condition

Objective To discuss the effects of Sry as an engrafting track of the bone marrow cells expanded under the different condition.Methods Using the methods of PCR,DOT Blot and in situ hybridization with specific primer or probe prepared according to the sequence of Y chromosome sex determining gene (Sry) of mice to track the engrafting state of the bone marrow cells expanded under the different condition. Results PCR and dot blot demonstrated that all of the female mouse transplanted with expanded or uncultured hematopoietic cells of the male had similar engraftment state. But the in situ hybridization showed that the mice transplanted with stromal cell supported expanded hematopoietic cells had more favourable condition. Conclusions Sry can be used as an index to determine the grafting results during hematopoietic stem cells transplantation. The results also suggest that the ex vivo hematopoietic cells expansion supported by bone marrow stromal cells have more favourable engrafting capacity.