间充质来源干细胞移植后注射粒细胞集落刺激因子可重建经致死剂量照射的BALB/c小鼠的造血功能
[Transplantation of mesenchymal derived stem cells followed by G-CSF injection can reconstitute hematopoiesis of lethally irradiated BALB/c mice].
作者信息
Hu Ying, Ma Li, Ma Guan-jie, Jiang Xue-ying, Zhao Chun-hua
机构信息
State Key Lab of Experimental Hematology, Institute of Hematology and Blood Diseases Hospital, CAMS, PUMC, Tianjin 300020, China.
出版信息
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2002 Feb;24(1):20-4.
OBJECTIVE
To explore the hematopoietic reconstitution potential of mesenchymal derived stem like cells.
METHODS
We transplanted bone marrow mesenchymal derived stem like cells into lethally irradiated BALB/c mice. Hematopoietic cells were derived from the non-adherent bone marrow cells 24 hours after initial culture while murine mesenchymal derived stem like cells from bone marrow of donor mice were cultured for 10 days before the transplantation.
RESULTS
All mice of group 1 and 3 died in 7-8 days post irradiation following transplantation, while all the mice from group 2 and 4 survived. The time course of hematopoietic reconstitution was then observed. The peripheral blood and bone marrow cell count recovered in the MSC + G-CSF transplanted group and the BM transplanted group after 3 weeks. Interestingly, CFU-GM number in the MSC + G-CSF transplanted group increased significantly after 2 weeks and even more than that in the BM transplanted group after 3 weeks while as CFU-GM colony dropped 2 weeks after in the BM transplanted group. Spleen colony (CFU-S) number and size of the MSC + G-CSF transplanted group was significantly greater than the BM transplanted group. Furthermore, PCR analysis was performed using peripheral blood cells to determine if any male-derived cells were present. No male-derived cells were found in any of the mice from group 1 and 3. Y-chromosome-specific src gene was found to be dominant in the MSC + G-CSF transplanted group and the BM transplanted group by week 4 post transplantation. In addition, we demonstrated that induction with G-CSF lead to CFU-GM colony formation from MSC compartment in vitro.
CONCLUSION
These results indicate that under stimulation of G-CSF, mesenchymal derived stem like cells might differentiate into hematopoietic primitive stem cells in vivo and have the capacity to re-establish hematopoiesis in lethally irradiated mice. This study should provide an alternative transplantation treatment for malignancy.
目的
探讨间充质来源的类干细胞的造血重建潜能。
方法
将骨髓间充质来源的类干细胞移植到经致死剂量照射的BALB/c小鼠体内。初始培养24小时后,非贴壁骨髓细胞用于获取造血细胞,而供体小鼠骨髓来源的鼠间充质来源的类干细胞在移植前培养10天。
结果
移植后,第1组和第3组的所有小鼠在照射后7 - 8天死亡,而第2组和第4组的所有小鼠均存活。随后观察造血重建的时间进程。3周后,间充质干细胞+粒细胞集落刺激因子(G-CSF)移植组和骨髓移植组的外周血和骨髓细胞计数恢复。有趣的是,间充质干细胞+G-CSF移植组的集落形成单位-粒细胞巨噬细胞(CFU-GM)数量在2周后显著增加,3周后甚至超过骨髓移植组,而骨髓移植组的CFU-GM集落在2周后下降。间充质干细胞+G-CSF移植组的脾集落(CFU-S)数量和大小显著大于骨髓移植组。此外,利用外周血细胞进行聚合酶链反应(PCR)分析,以确定是否存在任何雄性来源的细胞。在第1组和第3组的任何小鼠中均未发现雄性来源的细胞。移植后第4周,在间充质干细胞+G-CSF移植组和骨髓移植组中,Y染色体特异性src基因占主导地位。此外,我们证明在体外,G-CSF诱导可导致间充质干细胞区室形成CFU-GM集落。
结论
这些结果表明,在G-CSF刺激下,间充质来源的类干细胞可能在体内分化为造血原始干细胞,并具有在经致死剂量照射的小鼠中重建造血的能力。本研究应为恶性肿瘤提供一种替代性的移植治疗方法。