Type of Serum as a Cell Culture Supplement Influences Regulation of MicroRNA Expression in Breast MDA-MB-231 Cancer Cells

Nihad Al-Othman, Hana Hammad, Mamoun Ahram


Background: 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
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.


Charcoal-stripped fetal bovine serum, FBS, PCR array, 5α-dihydrotestosterone, Androgen receptor, Microrna, Breast cancer.

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