Synchrony in Human, Mouse and Bacterial Cell Cultures: A Comparison

Helmstetter, Charles E.; Thornton, Maureen; Romero, Ana; Eward, Kathryn Leigh

doi:10.4161/cc.2.1.185

Cited by 40 publications

(41 citation statements)

References 11 publications

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“…Recently, a new synchronization method in which cells are grown on a a A and B refer to the two basic methodologies described in Fig. 1 special device (baby-machine) and efficiently collected at the beginning of the cell cycle (Helmstetter et al 2003;Thornton et al 2002) was applied to this field. Use of this method achieved high temporal resolution with no perturbation of the cell cycle (Farkash-Amar et al 2008).…”

Section: Methodsmentioning

confidence: 99%

Genome-wide analysis of the replication program in mammals

Farkash-Amar¹,

Simon²

2009

Chromosome Res

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Microarray technology has facilitated the research of eukaryotic DNA replication on a genomewide scale. Recent studies have shed light on the association between time of replication and chromosome structure, on the organization principles of the replication program, and on the correlation between replication timing and transcription. In this review, we summarize various genomic measurement approaches and the biological insights achieved through applying them in the study of the mammalian replication program.

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Section: Methodsmentioning

confidence: 99%

Genome-wide analysis of the replication program in mammals

Farkash-Amar¹,

Simon²

2009

Chromosome Res

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“…Populations were synchronized using the membrane elution technique developed by Helmstetter (1969) and Helmstetter et al (2003). Briefly, cells were attached to a 0.22 mm pore diameter nitrocellulose membrane.…”

Section: Methodsmentioning

confidence: 99%

“…coli B/r A cultures grown in a minimal glucose medium were synchronized according to the membrane-elution technique developed by Helmstetter et al (2003). We adopted this technique to isolate cell cycle-related changes in transcript levels and to avoid changes in the gene expression profiles caused by temperature shifts or nutrient starvation techniques.…”

Section: E Coli Synchronous Cultures and Transcript Level Changesmentioning

confidence: 99%

Cell cycle progression in Escherichia coli B/r affects transcription of certain genes: Implications for synthetic genome design

Echtenkamp

Wilson

Shuler

2008

Biotech & Bioengineering

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We propose that transcript levels for some genes are affected by the bacterial cell division cycle and this may be an important factor to consider when designing synthetic bacterial genomes. To test this hypothesis, transcript levels of 58 genes in Escherichia coli B/r A were determined at five times during the cell division cycle. A two-step ANOVA technique was used to analyze data from custom oligonucleotide microarrays containing genes involved in important cellular processes including central metabolism, macromolecular synthesis, and transport and secretion. Consistent with results previously found in Caulobacter, approximately 17% of the transcript levels were cell cycle dependent. Cell cycle regulation can be divided into two classes: genes displaying increased transcript concentrations following gene replication and genes displaying an increased transcript concentration prior to replication initiation. Transcripts levels for hns, uspA, and zwf were affected by the cell division cycle, but did not fit well into either class. These results indicate that transcription of a significant fraction of the genome is affected by replication cycle progression. The results also show that both physical gene position and the physiological function of a gene affect when it is transcribed. In addition to the simple association with replication fork progression, other phenomena must be occurring to account for some of our observations. In conclusion, gene position, with regard to the C period, and gene function are important factors to incorporate into design criteria for synthetic bacterial genomes.

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“…The released newborn cells have been found to grow synchronously with no overt evidence of growth disturbances. Although this technique has been shown to function well (e.g., Cooper 2002;Helmstetter et al 2003;Eward et al 2004a, b;Grover et al 2004), in all instances the newborn cells were collected during the first few generations of growth after transfer from batch to immobilized growth.…”

Section: Introductionmentioning

confidence: 99%

Technology for cell cycle research with unstressed steady-state cultures

et al. 2006

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A culture system for performing cell cycle analyses on cells in undisturbed steady-state populations was designed and tested. In this system, newborn cells are shed continuously from an immobilized, perfused culture rotating about the horizontal axis. As a result of this arrangement, the number of newborn cells released into the effluent medium each generation is identical to the number of cells residing in the immobilized population, indicating that one of the two new daughter cells is shed at each cell division. Thus, the immobilized cells constitute a continuous, steady-state culture because the concentrations, locations and microenvironments of the cells in the culture vessel do not vary with time. In tests with mouse L1210 lymphocytic leukemia cells, about 10 8 newborn cells were produced per day. This new culture system enables a multiplicity of cell cycle analyses on large numbers of cells assured to be from populations in steady-state growth.

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Synchrony in Human, Mouse and Bacterial Cell Cultures: A Comparison

Cited by 40 publications

References 11 publications

Genome-wide analysis of the replication program in mammals

Genome-wide analysis of the replication program in mammals

Cell cycle progression in Escherichia coli B/r affects transcription of certain genes: Implications for synthetic genome design

Technology for cell cycle research with unstressed steady-state cultures

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