Tong Zhu, Gu Yongquan, Zhang Jian, Li Jianxin, Zhang Shuwen, Wang Zhonggao
Department of Vascular Surgery, Institute of Vascular Surgery, Xuanwu Hospital, Capital University of Medical Sciences, Beijing 100053, PR China.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2008 Oct;22(10):1218-21.
To observe the changes in the number and function of bone marrow-derived endothelial progenitor cells (EPCs) after bone-marrow stimulation, and to investigate the possible mechanism of improving ischemic limb disease after bone-marrow stimulation through autologoue bone-marrow stem cell implantation.
Twelve male Lewis rats, weighing 200-250 g, were classified into the bone marrow stimulation group (n = 6) and the control group (n = 6). In the stimulation group, the bone marrow of each rat was stimulated by injection of recombinant human granulocyte-macrophage colony-stimulatory factor. Mononuclear cells were harvested from bone marrow and cultured in EBM-2 medium. After 7-day culture, EPCs were stained by 1, 1-dioctadecyl-3, 3, 3, 3-tetramethylindocarbocyanine-labeled acetylated low density lipoprotein/fluorescein isothiocyanate-ulex europaeus agglutinin 1, and the double positive cells were counted by the fluorescent microscope. The adhesive ability of EPCs was determined by counting the number of re-cultured EPCs. The unilateral ischemia hindlimb model was made with 12 Lewis rats. Three days later, EPCs were transplanted into the ischemic tissues. According to different sources of EPCs, the 12 rats were divided into 2 groups: the stimulation group (n=6) and the control group (n = 6). At 3 weeks after EPCs transplantation, the quantity of the collateral vascular was observed by digital subtraction angiography (DSA).
After 7-day culture, the number of EPCs in the stimulation and control groups was (145.2 +/- 37.0)/HP and (95.2 +/- 39.4)/HP, respectively, and there was significant difference between the two groups (P < 0.05). Meanwhile, the number of adhesive EPCs in the stimulation and control groups was (21.8 +/- 4.3)/HP and (15.0 +/- 5.2)/HP, respectively, and the difference between the two groups was significant (P < 0.05). At 3 weeks after the EPCs implantation, the number of the collateral vascular was significantly larger in the stimulation group (4.2 +/- 1.2) compared with the control group (2.7 +/- 0.8), (P < 0.05).
Bone marrow stimulation increases the number of EPCs and improves the function concurrently, which may be the reason why autologoue bone-marrow stem cell implantation improves the curative effect of ischemic limb diseases after bone-marrow stimulation.
观察骨髓刺激后骨髓源性内皮祖细胞（EPCs）数量及功能的变化，探讨自体骨髓干细胞移植经骨髓刺激后改善肢体缺血性疾病的可能机制。
将12只体重200 - 250 g的雄性Lewis大鼠分为骨髓刺激组（n = 6）和对照组（n = 6）。刺激组大鼠通过注射重组人粒细胞巨噬细胞集落刺激因子刺激骨髓。从骨髓中采集单核细胞并在EBM - 2培养基中培养。培养7天后，用1,1 - 二油酰基 - 3,3,3,3 - 四甲基吲哚碳菁标记的乙酰化低密度脂蛋白/异硫氰酸荧光素 - 荆豆凝集素1对EPCs进行染色，并用荧光显微镜对双阳性细胞进行计数。通过对重新培养的EPCs计数来测定其黏附能力。用12只Lewis大鼠制作单侧缺血后肢模型。3天后，将EPCs移植到缺血组织中。根据EPCs的不同来源，将12只大鼠分为2组：刺激组（n = 6）和对照组（n = 6）。EPCs移植后3周，通过数字减影血管造影（DSA）观察侧支血管数量。
培养7天后，刺激组和对照组EPCs数量分别为（145.2 ± 37.0）/HP和（95.2 ± 39.4）/HP，两组间差异有统计学意义（P < 0.05）。同时，刺激组和对照组黏附EPCs数量分别为（21.8 ± 4.3）/HP和（15.0 ± 5.2）/HP，两组间差异有统计学意义（P < 0.05）。EPCs植入后3周，刺激组侧支血管数量（4.2 ± 1.2）明显多于对照组（2.7 ± 0.8），（P < 0.05）。
骨髓刺激可增加EPCs数量并同时改善其功能，这可能是自体骨髓干细胞移植经骨髓刺激后提高肢体缺血性疾病疗效的原因。
