Department of Stereotactic and Functional Neurosurgery, University of Freiburg, Freiburg, Germany.
Transplantation. 2011 Feb 27;91(4):390-7. doi: 10.1097/TP.0b013e3182063083.
The hippocampal formation is a specific structure in the brain where neurogenesis occurs throughout adulthood and in which the neuronal cell loss causes various demential states. The main goal of this study was to verify whether fetal neural progenitor cells (NPCs) from transgenic rats expressing green fluorescent protein (GFP) retain the ability to differentiate into neuronal cells and to integrate into the hippocampal circuitry after transplantation.
NPCs were isolated from E14 (gestational age: 14 days postconception) transgenic-Lewis and wild-type Sprague-Dawley rat embryos. Wild-type and transgenic cells were expanded and induced to differentiate into a neuronal lineage in vitro. Immunocytochemical and electrophysiological analysis were performed in both groups. GFP-expressing cells were implanted into the hippocampus and recorded electrophysiologically 3 months thereafter. Immunohistochemical analysis confirmed neuronal differentiation, and the yield of neuronal cells was determined stereologically.
NPCs derived from wild-type and transgenic animals are similar regarding their ability to generate neuronal cells in vitro. Neuronal maturity was confirmed by immunocytochemistry and electrophysiology, with demonstration of voltage-gated ionic currents, firing activity, and spontaneous synaptic currents. GFP-NPCs were also able to differentiate into mature neurons after implantation into the hippocampus, where they formed functional synaptic contacts.
GFP-transgenic cells represent an important tool in transplantation studies. Herein, we demonstrate their ability to generate functional neurons both in vitro and in vivo conditions. Neurons derived from fetal NPCs were able to integrate into the normal hippocampal circuitry. The high yield of mature neurons generated render these cells important candidates for restorative approaches based on cell therapy.
海马结构是大脑中的一个特定结构，在整个成年期和神经元细胞丢失时都会发生神经发生，导致各种痴呆状态。本研究的主要目的是验证表达绿色荧光蛋白（GFP）的转基因大鼠的胎儿神经前体细胞（NPC）在移植后是否仍然具有分化为神经元细胞并整合到海马回路中的能力。
从 E14 （妊娠年龄：受孕后 14 天）的转基因-Lewis 和野生型 Sprague-Dawley 大鼠胚胎中分离 NPC。将野生型和转基因细胞进行扩增，并在体外诱导分化为神经元谱系。在两组中均进行免疫细胞化学和电生理分析。将 GFP 表达细胞植入海马体，并在 3 个月后进行电生理记录。免疫组织化学分析证实了神经元分化，并通过立体学确定了神经元细胞的产量。
源自野生型和转基因动物的 NPC 在体外生成神经元细胞的能力相似。通过免疫细胞化学和电生理学证实了神经元成熟，表现为电压门控离子电流、放电活动和自发突触电流。GFP-NPC 也能够在植入海马体后分化为成熟神经元，并在其中形成功能性突触接触。
GFP 转基因细胞是移植研究中的重要工具。在此，我们证明了它们在体外和体内条件下生成功能性神经元的能力。源自胎儿 NPC 的神经元能够整合到正常的海马回路中。生成的成熟神经元产量高，使这些细胞成为基于细胞治疗的恢复方法的重要候选者。
