Alameddine H, Dehaupas M, Louboutin J P, Hantaï D, Joubert Y, Sébille A, Fardeau M
INSERM U 153, Paris, France.
C R Seances Soc Biol Fil. 1994;188(5-6):553-73.
We have developed an experimental model to test the ability of cultured satellite cells to form new muscle fibers when grafted in an irreversibly injured muscle. Association of X-irradiation to autotransplantation reduced extensor digitorum longus muscles of adult rats to a cystic structure formed by a peripheral rim of surviving muscle fibers surrounding a central space devoid of myofibers. Grafting autologous satellite cells, multiplied and labelled in vitro, into this central space gave rise to new myotubes which developed and matured to form fully differentiated myofibers. Mecanophysiological recordings confirmed the improvement of functional parameters particularly a 4 fold increase of twitch and tetanic tensions in grafted muscles. Furthermore, we examined the role of old basal lamina in architectural organisation of the reconstituted muscle by electron microscopy and immunofluorescence using antibodies to laminin, fibronectin, type IV collagen and heparan sulfate proteoglycan. We observed the persistence of immunoreactivity to all components; anti-laminin antibodies for example, evidenced a clear fascicular organization of ghost basal lamina which are progressively repopulated by the grafted cells. We finally investigated in vivo proliferation of the grafted cells by an autoradiographic study of H3-thymidine incorporation in the regenerated fibers and demonstrate that grafted cells proliferate for, at least, 1 week after cell grafting.
我们已经建立了一个实验模型，以测试培养的卫星细胞移植到不可逆损伤的肌肉中时形成新肌纤维的能力。将X射线照射与自体移植相结合，可使成年大鼠的趾长伸肌减少为一个囊性结构，该结构由围绕中央无肌纤维空间的存活肌纤维外周边缘形成。将体外增殖并标记的自体卫星细胞移植到这个中央空间，会产生新的肌管，这些肌管发育并成熟，形成完全分化的肌纤维。机械生理记录证实了功能参数的改善，特别是移植肌肉中的抽搐和强直张力增加了4倍。此外，我们通过电子显微镜和免疫荧光，使用针对层粘连蛋白、纤连蛋白、IV型胶原和硫酸乙酰肝素蛋白聚糖的抗体，研究了旧基膜在重建肌肉结构组织中的作用。我们观察到对所有成分的免疫反应性持续存在；例如，抗层粘连蛋白抗体证明了幽灵基膜的清晰束状组织，这些基膜逐渐被移植细胞重新填充。我们最后通过对再生纤维中H3-胸腺嘧啶核苷掺入的放射自显影研究，研究了移植细胞在体内的增殖情况，并证明移植细胞在细胞移植后至少增殖1周。
