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原代培养中星形胶质细胞的可塑性:一种实验工具及需谨慎对待方法学的原因。

Plasticity of astrocytes in primary cultures: an experimental tool and a reason for methodological caution.

作者信息

Juurlink B H, Hertz L

出版信息

Dev Neurosci. 1985;7(5-6):263-77. doi: 10.1159/000112295.

Abstract

Astrocytes in primary cultures constitute an exceedingly useful preparation for studies of astroglial development and function. These cells, however, demonstrate a pronounced plasticity in their reactions to culturing conditions. Thus, species and spatiotemporal region of CNS chosen for source of cells, dissociation procedures used, cell density in culture, culture medium chosen, type and/or concentration of serum used (if any) and exposure to dibutyryl cyclic AMP (dBcAMP) may all markedly affect the epigenotype of the cells. This provides an experimental tool for studies of astroglial development and function, but is at the same time a reason for caution in the interpretation of data when such cultured cells are used as models of their in vivo counterparts. About 95% of the cells in cultures used by the authors are positive for astrocyte-specific markers (glial fibrillary acidic protein and glutamine synthetase), and the cells possess presumably astrocytic characteristics such as high potassium permeability. This latter characteristic may be drastically altered by minor changes in culturing conditions. It is likely that an overwhelming majority of the astrocytes in such cultures are protoplasmic in nature. Addition of dBcAMP to the culture medium results in a pronounced morphological and a more modest functional differentiation.

摘要

原代培养的星形胶质细胞是研究星形胶质细胞发育和功能极为有用的样本。然而,这些细胞对培养条件的反应表现出显著的可塑性。因此,用于获取细胞的中枢神经系统的物种、时空区域、所采用的解离程序、培养中的细胞密度、所选用的培养基、所用血清的类型和/或浓度(如果使用)以及暴露于二丁酰环磷腺苷(dBcAMP),都可能显著影响细胞的表观基因型。这为研究星形胶质细胞的发育和功能提供了一种实验工具,但同时也是在将此类培养细胞用作其体内对应物模型来解释数据时需要谨慎的一个原因。作者所使用的培养物中约95%的细胞对星形胶质细胞特异性标志物(胶质纤维酸性蛋白和谷氨酰胺合成酶)呈阳性,并且这些细胞具有诸如高钾通透性等推测的星形胶质细胞特征。后一种特征可能会因培养条件的微小变化而发生剧烈改变。在此类培养物中,绝大多数星形胶质细胞可能本质上是原浆性的。向培养基中添加dBcAMP会导致明显的形态学分化和较为适度的功能分化。

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