Astrocyte Cultures Mimicking Brain Astrocytes in Gene Expression, Signaling, Metabolism and K(+) Uptake and Showing Astrocytic Gene Expression Overlooked by Immunohistochemistry and In Situ Hybridization

Neurochemical Research
Leif HertzDan Song

Abstract

Based on differences in gene expression between cultured astrocytes and freshly isolated brain astrocytes it has been claimed that cultured astrocytes poorly reflect the characteristics of their in vivo counterparts. This paper shows that this is not the case with the cultures of mouse astrocytes we have used since 1978. The culture is prepared following guidelines provided by Drs. Monique Sensenbrenner and John Booher, with the difference that dibutyryl cyclic AMP is added to the culture medium from the beginning of the third week. This addition has only minor effects on glucose and glutamate metabolism, but it is crucial for effects by elevated K(+) concentrations and for Ca(2+) homeostasis, important aspects of astrocyte function. Work by Liang Peng and her colleagues has shown identity between not only gene expression but also drug-induced gene upregulations and editings in astrocytes cultured by this method and astrocytes freshly isolated from brains of drug-treated animals. Dr. Norenberg's laboratory has demonstrated identical upregulation of the cotransporter NKCC1 in ammonia-exposed astrocytes and rats with liver failure. Similarity between cultured and freshly isolated astrocytes has also been shown in metabolism, K(+)...Continue Reading

References

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Citations

May 17, 2017·Neurotoxicology·Fabiana GallandMarina Concli Leite
Mar 7, 2018·Artificial Cells, Nanomedicine, and Biotechnology·Hang-Soo ParkSunghoi Hong
Dec 20, 2018·Physiological Reviews·Gerald A Dienel
Jun 28, 2018·Frontiers in Molecular Neuroscience·Yu-Feng Wang, Vladimir Parpura

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Methods Mentioned

BETA
electrophoresis
fluorescence-activated cell sorting
PCR
transgenic
NMR

Software Mentioned

FACSDiva

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