Hattiangady Bharathi, Shetty Ashok K
Institute for Regenerative Medicine, Texas A&M Health Science Center Temple, TX, USA.
Curr Protoc Stem Cell Biol. 2011 Sep;Chapter 2:Unit2D.7. doi: 10.1002/9780470151808.sc02d07s18.
Neural stem cell (NSC) transplantation into the hippocampus could offer an alternative therapy to hippocampal resection in patients with drug-resistant chronic epilepsy, which afflicts ∼30% of mesial temporal lobe epilepsy (TLE) cases. Multipotent, self-renewing NSCs could be expanded from multiple regions of the developing and adult brain, human embryonic stem cells (hESCs), and induced pluripotent stem cells (iPSCs). However, to provide a comprehensive methodology involved in testing the efficacy of transplantation of NSCs in a rat model of chronic TLE, NSCs derived from the embryonic medial ganglionic eminence (MGE) are taken as an example in this unit. The topics comprise description of the required materials, reagents and equipment, and protocols for expanding MGE-NSCs in culture, generating chronically epileptic rats, the intrahippocampal grafting, post-grafting evaluation of the effects of NSC grafts on spontaneous recurrent seizures and cognitive impairments, analyses of the yield and the fate of graft-derived cells, and the effects of NSC grafts on the host hippocampus.
将神经干细胞(NSC)移植到海马体中,可能为耐药性慢性癫痫患者提供一种替代海马体切除术的治疗方法,约30%的内侧颞叶癫痫(TLE)病例受此疾病困扰。多能、自我更新的神经干细胞可从发育中和成体大脑的多个区域、人类胚胎干细胞(hESC)以及诱导多能干细胞(iPSC)中扩增得到。然而,为了提供一种全面的方法,用于在慢性颞叶癫痫大鼠模型中测试神经干细胞移植的疗效,本单元以源自胚胎内侧神经节隆起(MGE)的神经干细胞为例。主题包括所需材料、试剂和设备的描述,以及在培养中扩增MGE-NSC的方案、生成慢性癫痫大鼠、海马体内移植、移植后评估神经干细胞移植对自发性复发性癫痫发作和认知障碍的影响、分析移植细胞的产量和命运,以及神经干细胞移植对宿主海马体的影响。