Neurospheres and glial cell cultures: immunocytochemistry for cell phenotyping.
Neurospheres and glial cell cultures: immunocytochemistry for cell phenotyping.
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神经球和神经胶质细胞培养物：用于细胞表型分析的免疫细胞化学。
DOI：
10.1007/978-1-62703-640-5_10
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发表时间：
2013
期刊：
影响因子：
--
通讯作者：
DelValle,Luis
中科院分区：
文献类型：
--
作者：
Struckhoff,AmandaParker;DelValle,Luis
细胞培养是一种重要的研究工具，是在受控系统中复制病理过程的一种方式。然而，单层脑源性细胞的培养提出了重大挑战，模糊了体外结果的保真度。经过几次传代后，神经胶质细胞和神经元细胞开始失去其形态学特征，最重要的是，失去其特异性细胞标志物和表型。近年来，神经祖细胞的发现和将其悬浮培养以保持其潜能同时仍保留分化成星形胶质细胞、少突胶质细胞和神经元的能力的方法学对神经科学和神经病理学领域做出了重大贡献。在脑中，祖细胞位于脑胶质细胞基质中，位于心室下区，并通过提供来源来替代因不同病理过程（例如损伤、遗传性疾病或疾病）而丢失或损坏的分化细胞，在大脑的稳态中发挥着重要作用。这些神经干细胞在传统上被认为具有有限或没有再生能力的器官中的发现为开发新的治疗方法打开了大门，其中包括细胞替代疗法。在这里，我们描述了从神经球神经祖细胞的培养和分化，以及使用免疫细胞化学对所得细胞进行表型分析。概述的免疫细胞学方法不限于神经球衍生培养物的分析，但也适用于原代神经胶质或细胞系衍生样品的细胞分型。
Cell cultures constitute an important tool for research as a way to reproduce pathological processes in a controlled system. However, the culture of brain-derived cells in monolayer presents significant challenges that obscure the fidelity of in vitro results. After a few number of passages, glial and neuronal cells begin to lose their morphological characteristics, and most importantly, their specific cellular markers and phenotype. In recent years, the discovery of Neural Progenitor Cells and the methodology to culture them in suspension maintaining their potentiality while still retaining the ability to differentiate into astrocytes, oligodendrocytes, and neurons have made significant contributions to the fields of neuroscience and neuropathology.In the brain, progenitor cells are located in the Germinal Matrix, in the subventricular zone and play an essential role in the homeostasis of the brain by providing the source to replace differentiated cells that have been lost or damaged by different pathological processes, such as injury, genetic conditions or disease. The discovery of these Neural Stem Cells in an organ traditionally thought to have limited or no regenerative capacity has open the door to the development of novel treatments, which include cell replacement therapy. Here we describe the culture and differentiation of neural progenitor cells from Neurospheres, and the phenotyping of the resulting cells using immunocytochemistry. The immunocytological methods outlined are not restricted to the analysis of Neurosphere-derived cultures but are also applicable for cell typing of primary glial or cell line-derived samples.