G-CSF动员外周血单个核细胞移植治疗大鼠缺血缺氧性脑病

Transplantation of Peripheral Blood Mononuclear Cells After GCSF Treatment for Hypoxia-Ischemia Therapy in Rats

目的:应用人外周血单个核细胞移植治疗新生大鼠缺血缺氧性脑病,探讨干细胞移植治疗神经系统损伤性疾病的有效性和安全性。方法:7 d龄SD大鼠随机分为3组(n=12),即正常组、手术组、细胞移植组;在征得同意的情况下利用粒细胞集落刺激因子(G-CSF)对捐献者的外周血进行动员,采用血细胞分离仪分离外周血单个核细胞,并进行鉴定及荧光标记;通过尾静脉注射的方法将标记细胞植入经免疫抑制剂处理的大鼠体内,利用HE染色法观察模型建立后大鼠大脑的损伤情况,利用荧光显微镜观察移植细胞在宿主体内的存活、迁移及分化情况,利用斜板实验明确细胞移植对宿主神经功能损伤情况的影响。结果:移植细胞可在宿主脑内存活,向损伤部位迁移,细胞移植可显著改善缺血缺氧引发的大脑功能损伤;细胞移植后,动物未现不良反应。结论:外周血单个核细胞移植治疗中枢系统损伤性疾病具有较高的安全性与有效性,有望成为一种临床治疗方案。

Objective： To observe the effects of cells transplantation on brain damage, we have witnessed an explosion in a novel therapeutic approach for hypoxia-ischemia（HI） in neonatal rat using the transplantation of human peripheral blood mononuclear cells（PB-MNC） from donors. Methods： 7-day-old SD rats were randomly divided into 3 groups（n=12）： control group, untreated HI group, and transplantation treated group. PB-MNC were isolated from G-CSF treated healthy after their approval using a CS-3000 blood cell separation machine. Then, the isolated cells were labeled with CM-Dil and transplanted into HI rats by tail-vein injection after surgery. Before transplantation, the rats were treated with immune-suppressive agent to promote donor cell survival. We observed the brain damage by HE staining, the survival, differentiation and migration of PB-MNC by fluorescent microscope, examined motor function of rats by tihboard tests. Results： PB-MNC could survive in the brain of hosts and locate in the damage area. In addition, compared with untreated HI rats, the HI rats received PB-MNC showed a reduction in motor function impairment, lesion volume and apoptosis ratio of neural cells. No adverse effect was observed in all animals after cell transplantation. Conclusion： Our results implied that transplantation of PB-MNC was of great safety and availability which may be a feasible candidate therapy method in clinic.