骨髓腔输注造血干/祖细胞在致敏模型应用的实验研究

Transfusion of Hematopoietic Stem/Progenitor Cells into Marrow Cavity in Sensitized Mouse Model

本研究旨在致敏模型中通过骨髓腔输注异基因造血干/祖细胞方法探讨致敏移植的新策略。应用脾细胞输注方法建立的致敏BABL/c小鼠作为致敏模型,取正常BALB/c小鼠作为非致敏受者,通过骨髓腔输注同种异基因造血干/祖细胞,观察移植后受者的生存及血象恢复情况。同时分离致敏及非致敏小鼠的血清,应用补体依赖细胞毒性反应方法检查血清抗体对异基因造血干/祖细胞损伤的影响。结果显示:非致敏受鼠经骨髓腔输注造血干/祖细胞后能长期存活,血象随时间推移而逐渐恢复正常。而致敏移植受鼠中有1只小鼠于移植前由于麻醉意外而死亡,其余90%致敏受鼠(9/10)经骨髓腔注射骨髓细胞后于2周内死亡,血象随时间推移而逐渐下降。取濒死的致敏受鼠组织行病理切片检查,证实致敏受鼠死于骨髓衰竭。补体依赖细胞毒性反应实验结果显示,非致敏组和致敏组的造血干/祖细胞死亡细胞百分比分别为(7.80±1.93)%和(50.80±3.12)%,两组差异有明显统计学意义(p<0.05),提示致敏血清对异基因造血干/祖细胞具有损伤作用。结论:通过骨髓腔输注造血干/祖细胞的方法并不能促进异基因供者细胞在致敏受者体内植入。

The study was aimed to investigate the strategy of transfusion of allogeneic hematopoietic stem/progenitor cells（HS/PC） into marrow cavity of mouse model in sensitized transplantation.A sensitized BALB/c mouse model was established by repeated transfusion of allogeneic spleen cells.The normal BALB/c mice were used as non-sensitized controls.The non-sensitized or sensitized recipients were transplanted by transfusion of allogeneic HS/PCs into bone marrow cavity.The survival rate and hematopoietic recovery were monitored.Moreover,non-sensitized and sensitized sera were obtained and incubated with allogeneic HS/PC respectively,the percentage of dead cells was calculated using complement-dependent cytotoxicity（CDC） tests.The results showed that non-sensitized recipients got long-term survival after the transfusion of HS/PC into marrow cavity,and the hematopoietic recovery increased along with time.However,among the sensitized recipients,one mouse died of anesthetic accident,the other 9 mice（9/10） died within 2 weeks after the transfusion of HS/PC in marrow cavity,and the hematopoietic recovery declined along with time.Histopathologic analysis demonstrated that the sensitized recipients died of bone marrow failure.The results of CDC tests showed that the percentage of dead cells in non-sensitized and sensitized group was 7.80±1.93% and 50.80±3.12%,respectively,and the differences were statistically significant（p〈0.05）,indicating sensitized sera were capable of impairing allogeneic HS/PC.It is concluded that the strategy of the marrow cavity transfusion of HS/PC can not enhance engraftment of allogeneic donor cells in sensitized recipients.