Elsevier

Brain Research Protocols

Volume 15, Issue 3, September 2005, Pages 115-118
Brain Research Protocols

Protocol
Reanalysis of the protocol for in vitro synchronization of mammalian astrocytic cultures by serum deprivation

https://doi.org/10.1016/j.brainresprot.2005.05.002Get rights and content

Abstract

Serum starvation of astrocytes for a period of time followed by refeeding has been proposed as a method to produce synchronized astrocytes. Here, it is proposed that the method neither synchronizes cells nor satisfies rigorous criteria for cell synchronization. The proposed non-selective, whole-culture synchronization protocol cannot, in theory, synchronize cells. The cells produced by the proposed serum starvation/refeeding protocol do not reflect the properties of any particular cell during the cell cycle. Cells produced by the published protocol will have a wide distribution of cell sizes, and therefore, the cells produced by starvation/refeeding will not model cells of any specific age during the division cycle. Thus, the proposed protocol will not produce a synchronized culture.

Section snippets

Commentary

It has been proposed that it is possible to synchronize astrocytes obtained from newborn rat brains by whole-culture treatment of cells [2]. The proposed method was to first starve cells in low serum, then refeed the cells with serum to obtain a synchronized culture. There are two parts to the detailed protocol: (1) proliferation of astrocytes under optimal conditions in vitro until the cells reach the desired confluence; and (2) synchronization of cultures by serum removal and proposed arrest

Discussion

The interest in the study of events during the cell cycle has led to numerous proposals for methods to synchronize cells. Here, one published method is analyzed. It is concluded that the published method neither presented evidence that the cells are synchronized nor considered theoretical arguments that the whole-culture method used–serum starvation followed by refeeding serum–does not, and cannot, synchronize cells [12].

Lest it be thought that it is proposed here that cell synchronization is

Acknowledgments

This work was supported by Grant MCB-0323346 from the National Science Foundation. Additional support for this research came (in part) from the National Institutes of Health through the University of Michigan's Cancer Center Support Grant (5 P30 CA46592).

References (21)

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