When performing cell culture, fetal bovine serum (FBS) is widely used to supplement in culture media. However, FBS may also contain compounds such as steroid hormones that can mask the effect of exogenous hormones under investigation. Processing of FBS with charcoal is a routine practice to remove these molecules; some of which may regulate the expression of microRNAs (miRNAs).

Aim:

To analyze the effect of the charcoal stripping of FBS when investigating regulation of miRNA expression by the steroid hormone, 5α-dihydrotestosterone (DHT) in breast cancer cells.

Methods:

Breast MDA-MB-231 cancer cells were cultured in media supplemented with either charcoal-stripped FBS (CS-FBS) or standard FBS. Cells were treated with 100 nM DHT for three days. Cell morphology was assessed. In addition, the expression of miRNAs was investigated using PCR arrays.

Results:

Alterations in cell morphology are serum-type independent. The basal expression of 13 miRNAs was slightly altered in cells cultured in CS-FBS versus standard FBS. Treatment of cells with DHT alters the expression of only four miRNA when grown in media supplemented with standard FBS. On the other hand, DHT alters the expression of 35 miRNAs when cells were cultured in media supplemented with CS-FBS.

Conclusion:

The type of serum influences miRNA expression in cultured cells and unmasks differences in miRNA expression induced by DHT.

Title

JORDAN MEDICAL JOURNAL

Publisher

The University of Jordan

Publisher Country

Jordan

Indexing

Scopus

Impact Factor

None

Publication Type

Prtinted only

Volume

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Year

--

Pages

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