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Mol. Hum. Reprod. Advance Access published online on March 8, 2006
Molecular Human Reproduction, doi:10.1093/molehr/gal018
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© The Author 2006. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org
Received November 17, 2005
Accepted January 16, 2006

Article

In vitro culture significantly alters gene expression profiles and reduces differences between myometrial and fibroid smooth muscle cells

Marina Zaitseva 1 *, Beverley J. Vollenhoven 1, and Peter A.W. Rogers 1

1 Centre for Women’s Health Research, Monash University Department of Obstetrics and Gynaecology, Monash Institute of Medical Research, Clayton, Victoria, Australia

* To whom correspondence should be addressed.
Marina Zaitseva, E-mail: marina.zaitseva{at}med.monash.edu.au


  Abstract

Cultured myometrial (M) and fibroid (F) smooth muscle cells (SMCs) have been widely used as a model for the study of F growth. The aim of this study was to compare gene expression profiles using microarrays between six paired M and F tissues from hysterectomy specimens, as well as cells isolated from the same tissues and cultured for up to three passages. A total of 2055 genes were differentially expressed by ANOVA between all experimental groups. Among them, 128 genes were found to be statistically different between M and F tissues. More than 1100 genes were significantly changed between tissues and cultured cells, with 648 genes common between both M and F cells at P0 and P3. Expression profiles of six genes including estrogen receptor-{alpha} (ER{alpha}) and pro-gesterone receptor (PR) were also validated using real-time PCR. These data demonstrate that large changes occur in SMC gene expression in culture, reducing differences between M and F cells. They also show that ER{alpha} and PR levels are reduced in cells compared with whole tissue. These results indicate that although M and F cell cultures provide an important tool to study these tumours, in vitro studies must be carefully planned and evaluated to provide meaningful results.

Keywords: fibroids/gene expression/in vitro culture/ER{alpha}/PR.
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