Pollock Kenneth, Stroemer Paul, Patel Sara, Stevanato Lara, Hope Andrew, Miljan Erik, Dong Ziping, Hodges Helen, Price Jack, Sinden John D
ReNeuron Ltd., 10 Nugent Road, Surrey Research Park, Guildford, Surrey GU2 7AF, UK.
Exp Neurol. 2006 May;199(1):143-55. doi: 10.1016/j.expneurol.2005.12.011. Epub 2006 Feb 7.
Transplantation of neural stem cells into the brain is a novel approach to the treatment of chronic stroke disability. For clinical application, safety and efficacy of defined, stable cell lines produced under GMP conditions are required. To this end, a human neural stem cell line, CTX0E03, was derived from human somatic stem cells following genetic modification with a conditional immortalizing gene, c-mycER(TAM). This transgene generates a fusion protein that stimulates cell proliferation in the presence of a synthetic drug 4-hydroxy-tamoxifen (4-OHT). The cell line is clonal, expands rapidly in culture (doubling time 50-60 h) and has a normal human karyotype (46 XY). In the absence of growth factors and 4-OHT, the cells undergo growth arrest and differentiate into neurons and astrocytes. Transplantation of CTX0E03 in a rat model of stroke (MCAo) caused statistically significant improvements in both sensorimotor function and gross motor asymmetry at 6-12 weeks post-grafting. In addition, cell migration and long-term survival in vivo were not associated with significant cell proliferation. These data indicate that CTX0E03 has the appropriate biological and manufacturing characteristics necessary for development as a therapeutic cell line.
将神经干细胞移植到大脑中是治疗慢性中风残疾的一种新方法。对于临床应用，需要在药品生产质量管理规范（GMP）条件下生产的明确、稳定的细胞系的安全性和有效性。为此，一种人类神经干细胞系CTX0E03是通过用条件永生化基因c-mycER(TAM)对人类体细胞干细胞进行基因改造后获得的。这种转基因产生一种融合蛋白，在合成药物4-羟基他莫昔芬（4-OHT）存在的情况下刺激细胞增殖。该细胞系是克隆性的，在培养中迅速扩增（倍增时间为50-60小时），并且具有正常的人类核型（46 XY）。在没有生长因子和4-OHT的情况下，细胞会停止生长并分化为神经元和星形胶质细胞。在中风大鼠模型（大脑中动脉闭塞，MCAo）中移植CTX0E03，在移植后6-12周，感觉运动功能和总体运动不对称性均有统计学上的显著改善。此外，细胞在体内的迁移和长期存活与显著的细胞增殖无关。这些数据表明，CTX0E03具有作为治疗性细胞系开发所需的适当生物学和生产特性。
