Schiaffino S, Gorza L, Sartore S, Saggin L, Carli M
Exp Cell Res. 1986 Mar;163(1):211-20. doi: 10.1016/0014-4827(86)90574-4.
Hybridoma cell lines were obtained from the fusion of NS-O myeloma cells with spleen cells of mice immunized with bovine fetal skeletal myosin. A stable hybridoma clone, BF-G6, produced immunoglobulin G1 k antibodies reacting specifically with embryonic-type myosin heavy chains present in fetal but not in neonatal or adult human skeletal muscle, as determined by enzyme immunoassay and immunoblot analysis. Fetal but not adult skeletal muscle fibers were stained by this monoclonal antibody in indirect immunofluorescence assays; smooth muscle cells and cardiac muscle cells, as well as non-muscle cells were also unreactive. Solid tumors of infants and children were tested for reactivity with BF-G6 by immunofluorescence and immunoperoxidase staining. Embryonic myosin heavy chain was expressed in rhabdomyosarcomas but not in other types of tumor, except for Wilms' tumor. Rhabdomyosarcoma cells isolated from a bone marrow metastasis and grown in vitro for several months were also labelled by BF-G6. Embryonic myosin heavy chain can thus be used as a specific differentiation marker of normal and neoplastic skeletal muscle tissue.
杂交瘤细胞系是通过将NS-O骨髓瘤细胞与用牛胎儿骨骼肌肌球蛋白免疫的小鼠脾细胞融合而获得的。一个稳定的杂交瘤克隆BF-G6产生免疫球蛋白G1 k抗体，通过酶免疫测定和免疫印迹分析确定，该抗体与胎儿而非新生儿或成人人类骨骼肌中存在的胚胎型肌球蛋白重链特异性反应。在间接免疫荧光试验中，该单克隆抗体可对胎儿而非成人骨骼肌纤维进行染色；平滑肌细胞、心肌细胞以及非肌肉细胞也无反应。通过免疫荧光和免疫过氧化物酶染色检测婴儿和儿童实体瘤与BF-G6的反应性。胚胎肌球蛋白重链在横纹肌肉瘤中表达，但在其他类型的肿瘤中不表达，除了Wilms瘤。从骨髓转移灶分离并在体外培养数月的横纹肌肉瘤细胞也被BF-G6标记。因此，胚胎肌球蛋白重链可作为正常和肿瘤性骨骼肌组织的特异性分化标志物。
