TRANSPLANTED HUMAN NEURONS DERIVED FROM A TERATOCARCINOMA CELL-LINE (NTERA-2) MATURE, INTEGRATE, AND SURVIVE FOR OVER 1 YEAR IN THE NUDE-MOUSE BRAIN
TRANSPLANTED HUMAN NEURONS DERIVED FROM A TERATOCARCINOMA CELL-LINE (NTERA-2) MATURE, INTEGRATE, AND SURVIVE FOR OVER 1 YEAR IN THE NUDE-MOUSE BRAIN
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DOI：
10.1002/cne.903570410
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发表时间：
1995-07-10
影响因子：
2.5
通讯作者：
LEE, VMY
中科院分区：
文献类型：
--
作者：
KLEPPNER, SR;ROBINSON, KA;LEE, VMY
视黄酸（RA）诱导人畸胎瘤细胞系（NTera-2或NT 2）仅产生有丝分裂后神经元样（NT 2N）细胞，但NT 2N细胞在体外从未获得完全成熟的神经元表型。为了确定NT 2N细胞是否可以在体内成熟为成人神经元样细胞，将纯化的NT 2N细胞移植到成年和新生无胸腺小鼠的中枢神经系统（CNS）的不同区域，并通过光、共聚焦、免疫组织化学和免疫组织化学检查移植物。以及使用一组发育调节神经元多肽的抗体进行电子显微镜检查。NT 2 N移植物与内源性小鼠神经元区分开来，识别所选神经元多肽中的人或鼠特异性表位的抗体。在> 89%的移植物受体（N = 90）中鉴定出活的NT 2N细胞，并且一些移植物存活14个月。在植入的3周内，移植的NT 2N细胞重新延长了它们的突起，并且移植物的位置（例如，隔膜与新皮层）似乎决定了加工的程度。在移植后早期，移植的NT 2N细胞表达与其体外对应物相同的神经元多肽。然而，在植入后6周和4-6个月之间，移植的NT 2N细胞逐渐获得完全成熟的体内神经元的分子表型，如通过重神经丝亚基的最高度磷酸化的同种型的表达的显著增加和成年CNS tau的从头表达所证明的。值得注意的是，在新生小鼠和成年小鼠中，NT 2N移植物的过程和神经元多肽表达的延长的时间过程是相似的。虽然移植的NT 2N细胞形成突触样结构和精心制作的树突和轴突，这些轴突仍然无髓鞘。这些研究表明，移植的人NT 2N细胞的纯群体在体内获得完全成熟的神经元表型，并且这些细胞在小鼠CNS中整合并存活> 1年。这些人类神经元样细胞是一个有吸引力的模型系统，用于研究神经元的发育，极性和移植。(C)1995 Wiley-Liss，Inc.
Retinoic acid (RA) induces a human teratocarcinoma cell line (NTera-2 or NT2) to give rise exclusively to post-mitotic neuron-like (NT2N) cells, but NT2N cells never acquire a fully mature neuronal phenotype in vitro. To determine whether NT2N cells can mature into adult neuron-like cells in vivo, purified NT2N cells were grafted into different regions of the central nervous system (CNS) of adult and neonatal athymic mice, and the grafts were examined immunohistochemically by light, confocal, and electron microscopy using antibodies to a panel of developmentally regulated neuronal polypeptides.NT2N grafts were distinguished from endogenous mouse neurons with antibodies that recognize human or murine specific-epitopes in selected neuronal polypeptides. Viable NT2N cells were identified in > 89% of graft recipients (N = 90), and some grafts survived 14 months. Within 3 weeks of implantation, grafted NT2N cells re-extended their processes, and the location of the grafts (e.g., septum versus neocortex) appeared to determine the extent to which processes were elaborated. Within the early post-transplantation period, grafted NT2N cells expressed the same neuronal polypeptides as their in vitro counterparts. However, between 6 weeks and 4-6 months post-implantation, the grafted NT2N cells progressively acquired the molecular phenotype of fully mature in vivo neurons as evidenced by dramatically increased expression of the most highly phosphorylated isoforms of the heavy neurofilament subunit, and the de novo expression of adult CNS tau. Notably, the time course for the extension of processes and the expression of neuronal polypeptides by NT2N grafts was similar in neonatal and adult mice. Although grafted NT2N cells formed synapse-like structures and elaborated dendrites and axons, these axons remained unmyelinated. Finally, none of the transplanted NT2N cells reverted to a neoplastic state.These studies demonstrate that pure populations of grafted human NT2N cells acquire a fully mature neuronal phenotype in vivo, and that these cells integrate and survive for > 1 year post-implantation in the mouse CNS. These human neuron-like cells are an attractive model system for studies of neuronal development, polarity and transplantation. (C) 1995 Wiley-Liss, Inc.