Please use this identifier to cite or link to this item: http://hdl.handle.net/10397/26625
Title: Purity, cell viability, expression of GFAP and bystin in astrocytes cultured by different procedures
Authors: Du, F
Qian, ZM
Zhu, L
Wu, XM
Qian, C
Chan, R
Ke, Y
Keywords: Astrocytes
Bystin
Glial fibrillary acidic protein (GFAP)
Ischemia/reperfusion
Primary culture
Reactive astrocytes
Shaken once culture
Shaken twice culture
Subculture
Issue Date: 2010
Publisher: Wiley-Liss
Source: Journal of cellular biochemistry, 2010, v. 109, no. 1, p. 30-37 How to cite?
Journal: Journal of Cellular Biochemistry 
Abstract: Primary astrocyte cultures are the most commonly used in vitro model for neurobiological studies. We speculated that different protocols might induce differences not only in the percentage of astrocytes but also in their biological characteristics. In this study, we investigated the effects of four major protocols on the purity of astrocytes, cell viability, expression of glial fibrillary acidic protein (GFAP) and bystin of cultured astrocytes using MTT assay, immunocytochemical staining, and Western blot analysis. We demonstrated that the purity of astrocytes (98.9%) generated by the subculture (SC) procedure is significantly higher than those generated by primary culture (PC), shaken once culture (SK-1) or shaken twice culture (SK-2). We also showed that expressions of GFAP and bystin in astrocytes that are purified by the SK-2 or SK-1 procedures are significantly higher than those in astrocytes prepared by PC or SC. In addition, astrocytes cultured by SK-2 or SK-1 have a higher level of cell viabilities at most time points after ischemia compared with astrocytes cultured by PC or SC. These suggested that physical stimulation induced by "shaken" or culture operation might be able to activate astrocytes and implied that different procedures induce differences not only in the purity but also in the biological characteristics of astrocytes, such as the percentage of activated astrocytes, GFAP, and bystin expressions and responses to ischemia. A more detailed analysis about the effect of "culture protocol factor" on the biological characteristics of astrocytes is absolutely needed.
URI: http://hdl.handle.net/10397/26625
DOI: 10.1002/jcb.22375
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