Torrente Y, Tremblay J P, Pisati F, Belicchi M, Rossi B, Sironi M, Fortunato F, El Fahime M, D'Angelo M G, Caron N J, Constantin G, Paulin D, Scarlato G, Bresolin N
Instituto de Ricovero e Cura a Carattere Scientifico (IRCCS) Ospedale Maggiore Policlinico, 20122 Milan, Italy.
J Cell Biol. 2001 Jan 22;152(2):335-48. doi: 10.1083/jcb.152.2.335.
Duchenne muscular dystrophy is a lethal recessive disease characterized by widespread muscle damage throughout the body. This increases the difficulty of cell or gene therapy based on direct injections into muscles. One way to circumvent this obstacle would be to use circulating cells capable of homing to the sites of lesions. Here, we showed that stem cell antigen 1 (Sca-1), CD34 double-positive cells purified from the muscle tissues of newborn mice are multipotent in vitro and can undergo both myogenic and multimyeloid differentiation. These muscle-derived stem cells were isolated from newborn mice expressing the LacZ gene under the control of the muscle-specific desmin or troponin I promoter and injected into arterial circulation of the hindlimb of mdx mice. The ability of these cells to interact and firmly adhere to endothelium in mdx muscles microcirculation was demonstrated by intravital microscopy after an intraarterial injection. Donor Sca-1, CD34 muscle-derived stem cells were able to migrate from the circulation into host muscle tissues. Histochemical analysis showed colocalization of LacZ and dystrophin expression in all muscles of the injected hindlimb in all of five out of five 8-wk-old treated mdx mice. Their participation in the formation of muscle fibers was significantly increased by muscle damage done 48 h after their intraarterial injection, as indicated by the presence of 12% beta-galactosidase-positive fibers in muscle cross sections. Normal dystrophin transcripts detected enzymes in the muscles of the hind limb injected intraarterially by the mdx reverse transcription polymerase chain reaction method, which differentiates between normal and mdx message. Our results showed that the muscle-derived stem cells first attach to the capillaries of the muscles and then participate in regeneration after muscle damage.
杜兴氏肌营养不良症是一种致命的隐性疾病，其特征是全身广泛的肌肉损伤。这增加了基于直接注射到肌肉中的细胞或基因治疗的难度。规避这一障碍的一种方法是使用能够归巢到损伤部位的循环细胞。在这里，我们表明，从新生小鼠肌肉组织中纯化的干细胞抗原1（Sca-1）、CD34双阳性细胞在体外具有多能性，并且可以进行肌源性和多髓样分化。这些肌肉衍生的干细胞是从在肌肉特异性结蛋白或肌钙蛋白I启动子控制下表达LacZ基因的新生小鼠中分离出来的，并注射到mdx小鼠后肢的动脉循环中。动脉内注射后，通过活体显微镜观察证明了这些细胞与mdx肌肉微循环中的内皮细胞相互作用并牢固粘附的能力。供体Sca-1、CD34肌肉衍生的干细胞能够从循环迁移到宿主肌肉组织中。组织化学分析显示，在五只接受治疗的8周龄mdx小鼠中的所有五只小鼠的注射后肢的所有肌肉中，LacZ和抗肌萎缩蛋白表达共定位。在动脉内注射48小时后进行的肌肉损伤显著增加了它们对肌纤维形成的参与，如肌肉横切面上12%的β-半乳糖苷酶阳性纤维的存在所示。通过mdx逆转录聚合酶链反应方法在动脉内注射的后肢肌肉中检测到正常的抗肌萎缩蛋白转录本，该方法可区分正常和mdx信息。我们的结果表明，肌肉衍生的干细胞首先附着于肌肉的毛细血管，然后在肌肉损伤后参与再生。
