Tsupykov Oleg, Kyryk Vitaliy, Smozhanik Ekaterina, Rybachuk Oksana, Butenko Gennadii, Pivneva Tatyana, Skibo Galina
Bogomoletz Institute of Physiology, Kyiv, Ukraine; State Key Laboratory of Molecular and Cellular Biology, Kyiv, Ukraine; State Institute of Genetic and Regenerative Medicine, Kyiv, Ukraine.
J Neurosci Res. 2014 Aug;92(8):964-74. doi: 10.1002/jnr.23386. Epub 2014 Apr 22.
The adult CNS has a very limited capacity to regenerate neurons after insult. To overcome this limitation, the transplantation of neural progenitor cells (NPCs) has developed into a key strategy for neuronal replacement. This study assesses the long-term survival, migration, differentiation, and functional outcome of NPCs transplanted into the ischemic murine brain. Hippocampal neural progenitors were isolated from FVB-Cg-Tg(GFPU)5Nagy/J transgenic mice expressing green fluorescent protein (GFP). Syngeneic GFP-positive NPCs were stereotactically transplanted into the hippocampus of FVB mice following a transient global cerebral ischemia model. Behavioral tests revealed that ischemia/reperfusion induced spatial learning disturbances in the experimental animals. The NPC transplantation promoted cognitive function recovery after ischemic injury. To study the long-term fate of grafted GFP-positive NPCs in a host brain, immunohistochemical approaches were applied. Confocal microscopy revealed that grafted cells survived in the recipient tissue for 90 days following transplantation and differentiated into mature neurons with extensive dendritic trees and apparent spines. Immunoelectron microscopy confirmed the formation of synapses between the transplanted GFP-positive cells and host neurons that may be one of the factors underlying cognitive function recovery. Repair and functional recovery following brain damage represent a major challenge for current clinical and basic research. Our results provide insight into the therapeutic potential of transplanted hippocampal progenitor cells following ischemic brain injury.
成年中枢神经系统在受到损伤后再生神经元的能力非常有限。为克服这一限制，神经祖细胞（NPCs）移植已发展成为神经元替代的关键策略。本研究评估了移植到缺血性小鼠脑内的NPCs的长期存活、迁移、分化及功能结果。从表达绿色荧光蛋白（GFP）的FVB-Cg-Tg(GFPU)5Nagy/J转基因小鼠中分离出海马神经祖细胞。在短暂性全脑缺血模型后，将同基因GFP阳性NPCs立体定向移植到FVB小鼠的海马体中。行为测试显示，缺血/再灌注在实验动物中诱导了空间学习障碍。NPC移植促进了缺血性损伤后认知功能的恢复。为研究移植的GFP阳性NPCs在宿主脑中的长期命运，应用了免疫组织化学方法。共聚焦显微镜显示，移植后90天，移植细胞在受体组织中存活，并分化为具有广泛树突和明显棘突的成熟神经元。免疫电子显微镜证实了移植的GFP阳性细胞与宿主神经元之间形成了突触，这可能是认知功能恢复的潜在因素之一。脑损伤后的修复和功能恢复是当前临床和基础研究面临的重大挑战。我们的结果为缺血性脑损伤后移植海马祖细胞的治疗潜力提供了见解。
