Philips M F, Mattiasson G, Wieloch T, Björklund A, Johansson B B, Tomasevic G, Martínez-Serrano A, Lenzlinger P M, Sinson G, Grady M S, McIntosh T K
Department of Neurosurgery, University of Pennsylvania School of Medicine and Veterans Administration Medical Center, Philadelphia, USA.
J Neurosurg. 2001 May;94(5):765-74. doi: 10.3171/jns.2001.94.5.0765.
Immortalized neural progenitor cells derived from embryonic rat hippocampus (HiB5), were transduced ex vivo with the gene for mouse nerve growth factor (NGF) to secrete NGF (NGF-HiB5) at 2 ng/hr/10(5) cells in culture.
Fifty-nine male Wistar rats weighing 300 to 370 g each were anesthetized with 60 mg/kg sodium pentobarbital and subjected to lateral fluid-percussion brain injury of moderate severity (2.3-2.4 atm, 34 rats) or sham injury (25 rats). At 24 hours postinjury, 2 microl (150,000 cells/microl) of [3H]thymidine-labeled NGF-HiB5 cells were transplanted stereotactically into three individual sites in the cerebral cortex adjacent to the injury site (14 rats). Separate groups of brain-injured rats received nontransfected (naive [n])-HiB5 cells (12 animals) or cell suspension vehicle (eight animals). One week postinjury, animals underwent neurological evaluation for motor function and cognition (Morris water maze) and were killed for histological, autoradiographic, and immunocytochemical analysis. Viable HiB5 cell grafts were identified in all animals, together with reactive microglia and macrophages located throughout the periinjured parenchyma and grafts (OX-42 immunohistochemistry). Brain-injured animals transplanted with either NGF-HiB5 or n-HiB5 cells displayed significantly improved neuromotor function (p < 0.05) and spatial learning behavior (p < 0.005) compared with brain-injured animals receiving microinjections of vehicle alone. A significant reduction in hippocampal CA3 cell death was observed in brain-injured animals receiving transplants of NGF-HiB5 cells compared with those receiving n-HiB5 cells or vehicle (p < 0.025).
This study demonstrates that immortalized neural stem cells that have been retrovirally transduced to produce NGF can markedly improve cognitive and neuromotor function and rescue hippocampal CA3 neurons when transplanted into the injured brain during the acute posttraumatic period.
将源自胚胎大鼠海马体的永生化神经祖细胞（HiB5）在体外转导小鼠神经生长因子（NGF）基因，使其在培养中以每10⁵个细胞每小时2纳克的速度分泌NGF（NGF-HiB5）。
59只体重300至370克的雄性Wistar大鼠，用60毫克/千克戊巴比妥钠麻醉，然后接受中度严重程度的侧方液体冲击性脑损伤（2.3 - 2.4个大气压，34只大鼠）或假损伤（25只大鼠）。在损伤后24小时，将2微升（每微升150,000个细胞）的[³H]胸腺嘧啶标记的NGF-HiB5细胞立体定向移植到损伤部位附近大脑皮层的三个独立位点（14只大鼠）。单独的脑损伤大鼠组接受未转染的（未处理的[n]）-HiB5细胞（12只动物）或细胞悬液载体（8只动物）。损伤后一周，对动物进行运动功能和认知（莫里斯水迷宫）的神经学评估，然后处死进行组织学、放射自显影和免疫细胞化学分析。在所有动物中均鉴定出存活的HiB5细胞移植体，以及遍布损伤周围实质和移植体中的反应性小胶质细胞和巨噬细胞（OX-42免疫组织化学）。与仅接受微注射载体的脑损伤动物相比，移植了NGF-HiB5或未处理的HiB5细胞的脑损伤动物在神经运动功能（p < 0.05）和空间学习行为（p < 0.005）方面有显著改善。与接受未处理的HiB5细胞或载体的脑损伤动物相比，接受NGF-HiB5细胞移植的脑损伤动物海马CA3区细胞死亡明显减少（p < 0.025）。
本研究表明，经逆转录病毒转导产生NGF的永生化神经干细胞在创伤后急性期移植到受损大脑中时，可显著改善认知和神经运动功能，并挽救海马CA3神经元。
