Mikhaĭlov V M, Evtifeeva E V, Serikov V B, Perverzev A E, Karmanova A V, Zenin V V
Tsitologiia. 2006;48(5):410-7.
Two sets of experiments were carried out. The first one involved chimeric mice, obtained by intravenously injections of bone marrow derived cells taken from transgenic C57BL/6 mice, expressing GFP, to 5 Gy X-ray irradiated mdx or C57BL/6 mice. In 2 months M. quadriceps femoris of chimeric mice were destroyed by surgical clamp. Following the next 4-5 weeks, the same muscles were studied for the presence of GFP-positive striated muscle fibres. In the case of chimeric C57BL/6 mice GFP-positive striated muscle fibres were observed in 0.3 +/- 0.5 and in 0.2 +/- 0.3 % of destroyed muscle, and in lateral (control) muscle, consequently. In the case of chimeric mdx mice, positive results were observed in 1.7 +/- 0.4 and in 0.5 +/- 0.3 % of destroyed and control muscles, respectively. In the second set of experiments, the GFP-positive bone marrow cells were used for multiple intramuscular injections to M. quadriceps femoris of C57BL/6 or mdx mice in a dose of 2 x 10(5)-5 x 10(5) cells per mouse. Before injection, GFP-positive bone marrow cells were fractionated in a 63 % Percoll solution and then were exhausted from differentiated cells by magnetic manner using CD4, CD8, CD38, CD45R, CD119, Ly-6G, and F4/80 antibodies. After 2-3 weeks, as many as 0.15 +/- 0.40 and 0.1 +/- 0.2 % of GFP-positive muscle fibres were found in injected and control muscles of C57BL/6 mice, respectively. In the case of mdx mice, the frequency of GFP-positive striated muscle fibres was 2.0 +/- 0.8 and 1.2 +/- 0.6 % for injected and control muscles, respectively. A conclusion is made that bone marrow stem cells can take part in differentiation of mdx mouse muscles after their delivery by needle injections.
进行了两组实验。第一组实验涉及嵌合小鼠,通过将取自表达绿色荧光蛋白(GFP)的转基因C57BL/6小鼠的骨髓来源细胞静脉注射到经5 Gy X射线照射的mdx或C57BL/6小鼠体内获得。2个月后,用手术夹破坏嵌合小鼠的股四头肌。在接下来的4 - 5周后,研究同一块肌肉中是否存在GFP阳性的横纹肌纤维。对于嵌合C57BL/6小鼠,在被破坏的肌肉中观察到GFP阳性横纹肌纤维分别占0.3±0.5%和0.2±0.3%,相应地,在外侧(对照)肌肉中也有观察到。对于嵌合mdx小鼠,在被破坏和对照肌肉中观察到阳性结果分别占1.7±0.4%和0.5±0.3%。在第二组实验中,将GFP阳性骨髓细胞以每只小鼠2×10⁵ - 5×10⁵个细胞的剂量多次肌肉注射到C57BL/6或mdx小鼠的股四头肌中。注射前,GFP阳性骨髓细胞在63%的 Percoll溶液中进行分级分离,然后使用CD4、CD8、CD38、CD45R、CD119、Ly - 6G和F4/80抗体通过磁性方法去除分化细胞。2 - 3周后,在C57BL/6小鼠的注射肌肉和对照肌肉中分别发现多达0.15±0.40%和0.1±0.2%的GFP阳性肌纤维。对于mdx小鼠,注射肌肉和对照肌肉中GFP阳性横纹肌纤维的频率分别为2.0±0.8%和1.2±0.6%。得出的结论是,骨髓干细胞通过针注射递送后可参与mdx小鼠肌肉的分化。