Upadhya Dinesh, Hattiangady Bharathi, Shetty Geetha A, Zanirati Gabriele, Kodali Maheedhar, Shetty Ashok K
Institute for Regenerative Medicine, Texas A&M University Health Science Center College of Medicine, Temple, Texas.
Research Service, Olin E. Teague Veterans' Medical Center, Central Texas Veterans Health Care System, Temple, Texas.
Curr Protoc Stem Cell Biol. 2016 Aug 17;38:2D.7.1-2D.7.47. doi: 10.1002/cpsc.9.
Grafting of neural stem cells (NSCs) or GABA-ergic progenitor cells (GPCs) into the hippocampus could offer an alternative therapy to hippocampal resection in patients with drug-resistant chronic epilepsy, which afflicts >30% of 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 human induced pluripotent stem cells (hiPSCs). On the other hand, GPCs could be generated from the medial and lateral ganglionic eminences of the embryonic brain and from hESCs and hiPSCs. To provide comprehensive methodologies involved in testing the efficacy of transplantation of NSCs and GPCs in a rat model of chronic TLE, NSCs derived from the rat medial ganglionic eminence (MGE) and MGE-like GPCs derived from hiPSCs are taken as examples in this unit. The topics comprise description of the required materials, reagents and equipment, methods for obtaining rat MGE-NSCs and hiPSC-derived MGE-like GPCs in culture, generation of chronically epileptic rats, intrahippocampal grafting procedure, post-grafting evaluation of the effects of grafts on spontaneous recurrent seizures and cognitive and mood impairments, analyses of the yield and the fate of graft-derived cells, and the effects of grafts on the host hippocampus. © 2016 by John Wiley & Sons, Inc.
将神经干细胞(NSCs)或γ-氨基丁酸能祖细胞(GPCs)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,慢性癫痫困扰着超过30%的颞叶癫痫(TLE)病例。多能、自我更新的神经干细胞可以从发育中和成体大脑的多个区域、人类胚胎干细胞(hESCs)以及人类诱导多能干细胞(hiPSCs)中扩增得到。另一方面,γ-氨基丁酸能祖细胞可以从胚胎脑的内侧和外侧神经节隆起以及人类胚胎干细胞和人类诱导多能干细胞中产生。为了提供在慢性颞叶癫痫大鼠模型中测试神经干细胞和γ-氨基丁酸能祖细胞移植疗效所涉及的综合方法,本单元以源自大鼠内侧神经节隆起(MGE)的神经干细胞和源自人类诱导多能干细胞的类似MGE的γ-氨基丁酸能祖细胞为例。主题包括所需材料、试剂和设备的描述,在培养中获得大鼠MGE神经干细胞和人类诱导多能干细胞衍生的类似MGE的γ-氨基丁酸能祖细胞的方法,慢性癫痫大鼠的产生,海马体内移植程序,移植后对移植物对自发复发性癫痫发作以及认知和情绪障碍影响的评估,对移植来源细胞的产量和命运的分析,以及移植物对宿主海马体的影响。© 2016约翰威立国际出版公司。