| Characterization of Hippocampal Adult-borne Granule Cells in a Transient Cerebral Ischemia Model
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Author:
Date:
2021-01-20
[Abstract] Long-term consequences of stroke significantly impair the quality of life in a growing population of stroke survivors. Hippocampal adult neurogenesis has been hypothesized to play a role in the pathophysiology of cognitive and neuropsychiatric long-term sequelae of stroke. Reliable animal models of stroke are paramount to understanding their biomechanisms and to advancing therapeutic strategies. We present a detailed protocol of a transient cerebral ischemia model which does not cause direct ischemic damage in the hippocampus, allowing investigations into the pathophysiology of long-term neurocognitive deficits of stroke. Furthermore, we describe a protocol for obtaining acute hippocampal slices for the purpose of electrophysiological and morphological characterization of adult-borne ...
[摘要] [摘要]中风的长期后果严重损害了越来越多的中风幸存者的生活质量。假定海马成年神经发生在中风的认知和神经精神病学长期后遗症的病理生理学中起作用。可靠的中风动物模型对于理解其生物机制和推进治疗策略至关重要。我们提出 暂时性脑缺血模型的详细协议，该模型不会在海马体中引起直接缺血损伤，从而可以研究中风的长期神经认知缺陷的病理生理学。此外，我们描述了一种用于获取成人海马颗粒细胞的电生理学和形态学特征的急性海马切片的协议。还讨论了与从小细胞（例如未成熟的成年颗粒细胞）进行电生理记录有关的特殊性。本协议可以通过多模式研究（行为，形态结构，生化）加以补充，以期有希望促进研究和中风的长期后遗症的发展以及发现新的治疗机会。
[背景]中风是在发达国家发病率和死亡率的重要原因引起急性和延迟赤字。虽然针对缺血性中风恢复血流的干预策略在降低急性发病率和死亡率方面已经变得更加有效，但中风的长期后果（如中风后抑郁和中风后认知功能障碍和痴呆症）目前正在逃避临床治疗（Wang等。，2010; Loubinoux等人，2012; Mijajlovic ...
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| Integration of Human Induced Pluripotent Stem Cell (hiPSC)-Derived Neurons into Rat Brain
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Author:
Date:
2020-09-05
[Abstract] Human neuron transplantation offers novel opportunities for modeling human neurologic diseases and potentially replacement therapies. However, the complex structure of the human cerebral cortex, which is organized in six layers with tightly interconnected excitatory and inhibitory neuronal networks, presents significant challenges for in vivo transplantation techniques to obtain a balanced, functional and homeostatically stable neuronal network. Here, we present a protocol to introduce human induced pluripotent stem cell (hiPSC)-derived neural progenitors to rat brains. Using this approach, hiPSC-derived neurons structurally integrate into the rat forebrain, exhibit electrophysiological characteristics, including firing, excitatory and inhibitory synaptic activity, and establish ...
[摘要] [摘要] 人类神经元移植为建模人类神经系统疾病和潜在的替代疗法提供了新的机会。然而，人脑皮层的复杂结构分为六层，具有紧密互连的兴奋性和抑制性神经元网络，这对体内移植技术获得平衡，功能稳定和稳态稳定的神经元网络提出了重大挑战。在这里，我们提出了一项协议，将人类诱导的多能干细胞（hiPSC ）衍生的神经祖细胞引入大鼠脑。使用这种方法，hiPSC 衍生的神经元在结构上整合到大鼠前脑中，表现出电生理特性，包括放电，兴奋性和抑制性突触活性，并与宿主电路建立神经元连通性。
[背景] 人类大脑皮层是一个复杂的细胞镶嵌体，在不同的皮质层（I-VI）中包含多样化的神经元亚型，可建立轴突输出和树突状输入的特定模式，提供了皮质电路的基本底物（Rakic，2009; Lodato 等等人，2011； Lui 等人，2011）。特别地，兴奋性和抑制性神经传递的平衡对于适当的脑功能是必需的（Turrigiano和Nelson，2004）。人类诱导的多能干细胞（hiPSC ）可以在人类遗传背景下对人类神经系统疾病进行建模（Dolmetsch和Geschwind，2011； Brennand 等，2015； Vera和Studer，2015）。建立体外系统以将hiPSCs ...
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| Isolation of THY1+ Undifferentiated Spermatogonia from Mouse Postnatal Testes Using Magnetic-activated Cell Sorting (MACS)
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Author:
Date:
2016-12-20
[Abstract] In mammals, homeostasis of many tissues rely on a subpopulation of cells, referred to as stem cells, to sustain an appropriate number of undifferentiated and differentiated cells. Spermatogonial stem cells (SSCs) provide the fundamental cellular source for spermatogenesis and are responsible for the lifelong maintenance of the germline pool in testes throughout the reproductive lifespan of males. To gain insight into germline stem cell biology and develop strategies for infertility treatment, several germ cell isolation methods have been reported in order to acquire good quality and quantity of undifferentiated spermatogonia. Among them, magnetic-activated cell sorting (MACS) is an efficient cell isolation method that requires less time and less initial cell numbers to obtain an enriched ...
[摘要] 在哺乳动物中，许多组织的体内平衡依赖于称为干细胞的细胞亚群，以维持适量的未分化细胞和分化细胞。精原细胞干细胞（SSC）为精子发生提供了基本的细胞来源，并且负责终生维持雄性生殖期间睾丸中的种系池。为了获得对种系干细胞生物学的了解并制定不孕症治疗策略，已经报道了几种生殖细胞分离方法，以获得良好质量和数量的未分化精原细胞。其中，磁激活细胞分选（MACS）是一种有效的细胞分离方法，其需要较少的时间和较少的初始细胞数以使用抗原 - 抗体反应来获得富集的细胞群体。胸腺细胞抗原1（THY1，CD90.2）被认为是小鼠新生儿和成年睾丸中未分化精原细胞的表面标志物。在这里，我们描述了一个用于分离来自小鼠睾丸的一周龄THY1 + 细胞和四周龄THY1 + 细胞的方案。分离过程包括三个步骤：睾丸收集和单细胞悬浮，使用生物素缀合的抗THY1抗体的细胞标记和磁性细胞分离。注意，这种隔离方案应在五小时内完成，以最大限度地提高活细胞的质量和数量。
背景 在几个成人组织中观察到活性和静止干细胞的共存。静止，自我更新和分化之间的充分平衡对于维持适当数量的未分化干细胞是必要的，并且避免过早的干细胞耗尽许多组织的体内平衡（Tseng等人，2015; Wabik和Jones，2015; ...
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