骨髓间充质干细胞在致敏与非致敏BALB/c小鼠造血干细胞移植中的应用

Application of bone marrow mesenchymal stem cells in hematopoietic stem cell transplantation for sensitized and non-sensitized BALB/c mice

背景:造血干细胞移植对多种疾病具有治疗作用,但其取材不便,且细胞数量受年龄限制等原因,故而应用具有一定局限性。目的:探索骨髓间充质干细胞在致敏与非致敏BALB/c小鼠造血干细胞移植中的应用价值。方法:将BALB/c小鼠骨髓细胞在体外进行分离,采用贴壁培养的方法获得间充质干细胞,使用流式细胞仪对细胞表面的分子标记进行检测。应用异基因脾细胞输注方法建立致敏动物模型,用绿色荧光染料标记骨髓间充质干细胞,分别移植到致敏和非致敏的受体小鼠体内,并在移植后的不同时间点对间充质干细胞的归巢情况进行检测。对致敏BALB/c小鼠进行照射预处理,联合应用异基因骨髓细胞与同基因间充质干细胞移植,观察BALB/c小鼠的生存情况。结果与结论:移植48 h后,间充质干细胞在致敏受体和非致敏受体小鼠分别归巢于脾脏和骨髓。在造血干细胞的移植实验中,致敏BALB/c小鼠接受异基因骨髓细胞与同基因骨髓间充质干细胞联合移植,结果显示致敏BALB/c小鼠全部在移植后12-15 d死亡,生存的中位时间是14 d,而仅接受异基因骨髓细胞移植的致敏BALB/c小鼠的中位生存时间为13 d。说明细胞移植后在致敏受体内间充质干细胞主要归巢为脾脏和骨髓,联合应用间充质干细胞移植对异基因造血干/祖细胞植入致敏受体体内并没有起到有效的促进作用。

BACKGROUND：Hematopoietic stem cel transplantation has therapeutic effects on many diseases, but its application has some limitations, such as cel harvesting and age-limited number of cels. OBJECTIVE： To investigate the application value of bone marrow mesenchymal stem cels in hematopoietic stem cel transplantation for sensitized and non-sensitized BALB/c mice. METHODS： Bone marrow cels derived from BALB/c mice were isolated and culturedin vitro to harvest mesenchymal stem cels using adherent method. The cel surface markers were detected by flow cytometry. A murine model of sensitization was established by transfusion of alogeneic spleen cels. Mesenchymal stem cels labeled with green fluorescent dye were transplanted into non-sensitized and sensitized recipient mice, and the homing of mesenchymal stem cels in vivowas monitored at different time points post transplantation. Additionaly, under irradiation pretreatment, sensitized BALB/c mice under irradiation were subjected to combined transplantation of alogeneic bone marrow cels and syngeneic mesenchymal stem cels. Survival rate of BALB/c mice was monitored daily. RESULTS AND CONCLUSION： At 48 hours after transplantation, mesenchymal stem cels in sensitized and non-sensitized recipients were homing to the spleen and bone marrow, respectively. In the experiment of hematopoietic stem cel transplantation, the sensitized recipients died at 12-15 days after combined＆nbsp;transplantation, with a median of 14 days; however, the sensitized recipients only undergoing alogeneic bone marrow cel transplantation had a survival median of 13 days. These findings indicate that the transplanted mesenchymal stem cels in sensitized recipients are mainly homing to the spleen and bone marrow, but the combination transplantation cannot enhance the transplantation of alogeneic hematopoietic stem/progenitor cels in sensitized recipients.