Zhang Chen, Saatman Kathryn E, Royo Nicolas C, Soltesz Kristie M, Millard Marie, Schouten Joost W, Motta Melissa, Hoover Rachel C, McMillan Asenia, Watson Deborah J, Lee Virginia M-Y, Trojanowski John Q, McIntosh Tracy K
Traumatic Brain Injury Laboratory, Department of Neurosurgery, University of Pennsylvania, 105 Hayden Hall, 3320 Smith Walk, Philadelphia, PA 19104, USA.
J Neurotrauma. 2005 Dec;22(12):1456-74. doi: 10.1089/neu.2005.22.1456.
The NTera2 (NT2) cell line is a homogeneous population of cells, which, when treated in vitro with retinoic acid, terminally differentiate into postmitotic neuronal NT2N cells. Although NT2N neurons transplanted in the acute (24 h postinjury) period survive for up to 1 month following experimental traumatic brain injury (TBI), nothing is known of their ability to survive for longer periods or of their effects when engrafted during the chronic postinjury period. Adult male Sprague-Dawley rats (n = 348; 360-400 g) were initially anesthetized and subjected to severe lateral fluid-percussion (FP) brain injury or sham injury. At 1 month postinjury, only brain-injured animals showing severe neurobehavioral deficits received cryopreserved NT2N neurons stereotaxically transplanted into three sites in the peri-injured cortex (n = 18). Separate groups of similarly brain-injured rats received human fibroblast cells (n = 13) or cell suspension vehicle (n = 14). Sham-injured animals (no brain injury) served as controls and received NT2N transplants (n = 24). All animals received daily immunosuppression for three months. Behavioral testing was performed at 1, 4, 8, and 12 weeks post-transplantation, after which animals were sacrificed for histological analysis. Nissl staining and anti-human neuronal specific enolase (NSE) immunostaining revealed that NT2N neurons transplanted in the chronic post-injury period survived up to 12 weeks post-transplantation, extended processes into the host cortex and immunolabeled positively for synaptophysin. There were no statistical differences in cognitive or motor function among the transplanted brain-injured groups. Long-term graft survival suggests that NT2N neurons may be a viable source of neural cells for transplantation after TBI and also that these grafts can survive for a prolonged time and extend processes into the host cortex when transplanted in the chronic post-injury period following TBI.
NTera2（NT2）细胞系是一群同质细胞，当在体外用视黄酸处理时，它们会终末分化为有丝分裂后神经元NT2N细胞。尽管在实验性创伤性脑损伤（TBI）后的急性期（损伤后24小时）移植的NT2N神经元能存活长达1个月，但对于它们在更长时期内存活的能力以及在损伤后慢性期植入时的影响却一无所知。成年雄性Sprague-Dawley大鼠（n = 348；360 - 400克）最初接受麻醉，然后遭受严重的侧方液体冲击（FP）脑损伤或假损伤。在损伤后1个月，只有表现出严重神经行为缺陷的脑损伤动物接受立体定向移植到损伤周围皮质三个部位的冷冻保存的NT2N神经元（n = 18）。单独的几组类似脑损伤大鼠接受人成纤维细胞（n = 13）或细胞悬液载体（n = 14）。假损伤动物（无脑损伤）作为对照并接受NT2N移植（n = 24）。所有动物接受为期三个月的每日免疫抑制。在移植后1、4、8和12周进行行为测试，之后处死动物进行组织学分析。尼氏染色和抗人神经元特异性烯醇化酶（NSE）免疫染色显示，在损伤后慢性期移植的NT2N神经元在移植后存活长达12周，其突起延伸到宿主皮质，并对突触素呈阳性免疫标记。移植的脑损伤组之间在认知或运动功能方面没有统计学差异。长期移植存活表明，NT2N神经元可能是TBI后移植的可行神经细胞来源，并且这些移植物在TBI后的损伤后慢性期移植时可以长时间存活并将突起延伸到宿主皮质。
