Trotter J, Crang A J, Schachner M, Blakemore W F
Department of Neurobiology, University of Heidelberg, Germany.
Glia. 1993 Sep;9(1):25-40. doi: 10.1002/glia.440090105.
Immortalised lines of murine glial precursor cells expressing the neomycin resistance gene and a temperature-sensitive mutation of the SV 40 T oncogene were established from cultures of oligodendrocytes and precursor cells infected with a replication-incompetent, helper-free retrovirus. At the permissive temperature (33 degrees C), they could be continually propagated in vitro and cells were present expressing the 04 antigen specific for glial precursor cells and oligodendrocytes. At 38 degrees C, where the expression of the T antigen is down regulated, cell division largely ceased. During early passage in vitro, limited differentiation to a more mature phenotype, as evidenced by expression of GFAP and the oligodendrocyte marker 01 was observed at both 33 degrees C and 38 degrees C. When transplanted into demyelinating lesions in the spinal cords of adult rats early passages of the lines yielded myelin-forming oligodendrocytes and astrocytes. Cells from later passages of the lines although failing to synthesise myelin still associated specifically with the demyelinated axons. These experiments demonstrate the retention of physiological properties of these oncogene-carrying glial cells when transplanted in vivo and suggest that such immortalised populations can be used for the isolation of molecules regulating glial cell function.
利用无复制能力、无辅助病毒的逆转录病毒感染少突胶质细胞和前体细胞培养物,建立了表达新霉素抗性基因和SV40 T癌基因温度敏感突变的永生化小鼠神经胶质前体细胞系。在允许温度(33摄氏度)下,它们可在体外持续传代培养,且存在表达神经胶质前体细胞和少突胶质细胞特异性04抗原的细胞。在38摄氏度时,T抗原表达下调,细胞分裂基本停止。在体外早期传代过程中,在33摄氏度和38摄氏度时均观察到有限程度的向更成熟表型分化,表现为GFAP和少突胶质细胞标志物01的表达。当将该细胞系的早期传代细胞移植到成年大鼠脊髓的脱髓鞘损伤部位时,可产生形成髓鞘的少突胶质细胞和星形胶质细胞。该细胞系后期传代的细胞虽然不能合成髓鞘,但仍能特异性地与脱髓鞘轴突相关联。这些实验证明了这些携带癌基因的神经胶质细胞在体内移植时保留了生理特性,并表明这种永生化细胞群体可用于分离调节神经胶质细胞功能的分子。