Survival and integration of transplanted postmitotic human neurons following experimental brain injury in immunocompetent rats.
Survival and integration of transplanted postmitotic human neurons following experimental brain injury in immunocompetent rats.
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DOI：
10.3171/jns.1999.90.1.0116
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发表时间：
1999
影响因子：
4.1
通讯作者：
Matthew F. Philips;J. K. Muir;K. Saatman;R. Raghupathi;V. Lee;J. Trojanowski;Tracy K. McIntosh
中科院分区：
文献类型：
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作者：
Matthew F. Philips;J. K. Muir;K. Saatman;R. Raghupathi;V. Lee;J. Trojanowski;Tracy K. McIntosh
对象有关细胞同质性和存活性的限制不会影响神经元样 hNT 细胞，此类 hNT 细胞源自人畸胎癌细胞系 (Ntera2)，接触视黄酸可分化为有丝分裂后神经元。由于 NT2N 神经元在移植到裸鼠大脑中后存活时间超过 1 年，因此作者将这些细胞移植到侧向液体冲击性脑损伤后免疫功能正常的大鼠大脑中，以确定 NT2N 细胞移植物在创伤性脑损伤 (TBI) 受损皮质中的长期生存能力以及 NT2N 神经元对认知和运动缺陷的治疗效果。方法 72 只成年雄性 Sprague-Dawley 大鼠，每只体重在 340 至 370 g 之间，给予麻醉剂并进行中度侧面液体冲击脑损伤（46 只大鼠为 2.2-2.5 atm）或接受无 TBI 的手术（假手术，26 只大鼠）。损伤后24小时，将10(5)个NT2N细胞（24只受伤动物）或3微升媒介物（22只受伤动物和14只对照动物）立体定向植入受伤周围或对照大脑皮层。每周评估一次运动功能，并在使用 Morris 水迷宫范例评估其创伤后学习能力后 2 或 4 周处死所有动物。常规观察到，在移植后 2 周和 4 周时，活的 NT2N 移植物将人神经细胞粘附分子 (MOC-1) 免疫反应过程延伸到受伤周围的皮质中，但没有发现运动或认知功能有显着改善。通过巨噬细胞 (ED-1) 和小胶质细胞 (异凝集素 B4) 的免疫组织化学鉴定来评估两个时间点移植周围发现的炎症。结论 NT2N 细胞在免疫功能正常的大鼠受伤周围皮层中的长期存活和整合为研究人员提供了一个重要的细胞系统，可用于研究 TBI 后移植神经元的成熟、调节和神经突生长。
OBJECT Limitations regarding cell homogeneity and survivability do not affect neuronlike hNT cells, which are derived from a human teratocarcinoma cell line (Ntera2) that differentiates into postmitotic neurons with exposure to retinoic acid. Because NT2N neurons survive longer than 1 year after transplantation into nude mice brains, the authors grafted these cells into the brains of immunocompetent rats following lateral fluid-percussion brain injury to determine the long-term survivability of NT2N cell grafts in cortices damaged by traumatic brain injury (TBI) and the therapeutic effect of NT2N neurons on cognitive and motor deficits. METHODS Seventy-two adult male Sprague-Dawley rats, each weighing between 340 and 370 g, were given an anesthetic agent and subjected to lateral fluid percussion brain injury of moderate severity (2.2-2.5 atm in 46 rats) or to surgery without TBI (shamoperation, 26 rats). Twenty-four hours postinjury, 10(5) NT2N cells (24 injured animals) or 3 microl of vehicle (22 injured and 14 control animals) was stereotactically implanted into the periinjured or control cerebral cortex. Motor function was assessed at weekly intervals and all animals were killed at 2 or 4 weeks after their posttraumatic learning ability was assessed using a Morris water maze paradigm. Viable NT2N grafts were routinely observed to extend human neural cell adhesion molecule-(MOC-1)immunoreactive processes into the periinjured cortex at 2 and 4 weeks posttransplantation, although no significant improvement in motor or cognitive function was noted. Inflammation identified around the transplant at both time points was assessed by immunohistochemical identification of macrophages (ED-1) and microglia (isolectin B4). CONCLUSIONS Long-term survival and integration of NT2N cells in the periinjured cortex of immunocompetent rats provides the researcher with an important cellular system that can be used to study maturation, regulation, and neurite outgrowth of transplanted neurons following TBI.