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Table 1 Analysis of differential gene expression in HeLa triggered by different cultivation temperatures.

From: Novel approaches to render stable producer cell lines viable for the commercial manufacturing of rAAV-based gene therapy vectors

Name

Differential expression at

Mode of regulation

Microarray analysis

RT qPCR

Gene A

30°C

Up

3.2-fold

6.9-fold

Gene B

30°C

Up

2.2-fold

2.6-fold

Gene C

30°C

Up

3.3-fold

2.3-fold

miRNA A

32°C

Up

3.1-fold

-

miRNA B

32°C

Down

3.3-fold

-

miRNA C

32°C

Up

3.0-fold

-

  1. Cells were seeded at two different densities and cultivated at 37°C for two days. Subsequently, cells were incubated for another 6 hours at 30, 32, and 37°C, respectively, before mRNA was isolated from the cells. Microarray analysis (GeneChip® Human Exon 1.0 ST Array, Affymetrics) was performed to identify mRNAs differentially expressed more than 2-fold. Validation was done by RT qPCR analysis (EvaGreen® Mastermix, Biorad) and included controls of regulated and non-regulated mRNAs [12, 11, 1]. Differentially expressed miRNAs (>2-fold) were also identified by microarray analysis (GeneChip® miRNA 2.0 Array, Affymetrics). As validation is not yet completed, only an excerpt of the most promising miRNA candidates is shown.