Onifer S M, Whittemore S R, Holets V R
Miami Project to Cure Paralysis, Department of Neurological Surgery, University of Miami School of Medicine, Florida 33136.
Exp Neurol. 1993 Jul;122(1):130-42. doi: 10.1006/exnr.1993.1114.
A clonal, neuronally-differentiating cell line, RN33B, was previously developed by retroviral infection of neural tissue derived from embryonic Sprague-Dawley raphé nuclei with a retrovirus encoding the temperature-sensitive allele of SV40 large T-antigen. In the present study, RN33B cells were transplanted into two target areas of the raphé nuclei, the spinal cord and hippocampal formation, of adult allogeneic hosts. Prior to transplantation, RN33B cells were infected in vitro with a retroviral vector carrying the Escherichia coli lacZ reporter gene and were visualized in vivo using a beta-galactosidase immunohistochemical technique. RN33B cells were seen throughout the spinal cord and hippocampal formation of the adult hosts at 15 days post-transplantation. T-antigen-immunoreactive nuclei were detected where RN33B cells were observed, but in much greater numbers than beta-galactosidase-immunoreactive cells. Bipolar RN33B cells were found in the spinal cord grey matter. RN33B cells with multipolar morphologies were visualized in the hippocampal and subicular pyramidal cell layers, and also in the dentate gyrus granule cell and polymorph layers, while bipolar RN33B cells were seen in the remainder of the hippocampal formation. The results suggest that immortalized neural cell lines of CNS origin can differentiate in the adult CNS with their ultimate morphology being determined by local tissue signals. We speculate that endogenous neutrophins may significantly influence RN33B cell differentiation in vivo.
一种克隆的、具有神经元分化能力的细胞系RN33B，先前是通过用编码SV40大T抗原温度敏感等位基因的逆转录病毒感染源自胚胎斯普拉格-道利大鼠缝际核的神经组织而建立的。在本研究中，将RN33B细胞移植到成年同种异体宿主的缝际核的两个靶区域，即脊髓和海马结构。在移植前，将携带大肠杆菌lacZ报告基因的逆转录病毒载体在体外感染RN33B细胞，并使用β-半乳糖苷酶免疫组织化学技术在体内进行可视化观察。移植后15天，在成年宿主的整个脊髓和海马结构中都可见到RN33B细胞。在观察到RN33B细胞的地方检测到T抗原免疫反应性细胞核，但其数量比β-半乳糖苷酶免疫反应性细胞多得多。在脊髓灰质中发现了双极RN33B细胞。在海马和海马下锥体细胞层以及齿状回颗粒细胞层和多形层中可见到具有多极形态的RN33B细胞，而在海马结构的其余部分可见到双极RN33B细胞。结果表明，源自中枢神经系统的永生化神经细胞系可以在成年中枢神经系统中分化，其最终形态由局部组织信号决定。我们推测内源性神经营养因子可能在体内显著影响RN33B细胞的分化。
