Morphological comparison of ectopic and eutopic endometriotic stromal cells in culture

Nastaran Miyanmahaleh,1 Zeynab borandegi,2 Maryam shahhoseini,3,* Parvaneh afsharian,4 Ashraf moini,5

1. 1. Department of Molecular genetics, Faculty of Basic Sciences and Advanced Technologies in Biology, University of Science and Culture, ACECR, Tehran, Iran. 2. Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
2. 1. Department of Molecular genetics, Faculty of Basic Sciences and Advanced Technologies in Biology, University of Science and Culture, ACECR, Tehran, Iran. 2. Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
3. Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran , Iran
4. Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
5. 1. Department of Endocrinology and Female Infertility, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran. 2. Department of Obstetrics and Gynecology, Arash Womens Hospital, Tehran University of Medical Sciences, Tehran, Iran.

Abstract

Introduction

Endometriosis is a benign disease which is caused by presence of endometrial tissue including glandular and stromal cells in the extra-uterine environment. it has a prevalence of 10–15% among women of reproductive age and may cause severe chronic pelvic pain and infertility. the effects of endometriosis on womens quality of life can explain the importance of in-vitro study of this disease. it is known that there are differences between many biological features of eutopic and ectopic endometrial tissues such as invasive and adhesive behavior, growth, gene expression profile and epigenetics. in this study we compared microscopic phenotype, proliferation rate and the surface area of eutopic endometrial stromal cells (euescs) with ectopic endometrial stromal cells (eescs).

Methods

Samples were obtained from 2 patients with ovarian endometriosis (endometrioma) during the proliferative phase of the menstrual cycle at arash womens hospital. eescs and euescs were isolated and cultured in phenol red free dulbeccos modified eagles medium (dmem) with 10% fetal bovine serum (fbs). the cells were passaged to passage 3 in which the fluorescence activated cell sorting (facs) analysis showed more than 99% purity of stromal cells. then the morphology and the proliferation rate was assessed by an inverted microscope.

Results

Eescs were larger in size compared to euescs morphology and had lower proliferation rate than euescs. in addition, the cell density of eescs were significantly lower than euescs in equal culture conditions and seeding density at confluency of 90% in 6 well plates. the estimated surface area that each of the ectopic stromal cells occupied (48× 10 -4 mm2) was about 3 times more than the eutopic stromal cells (14× 10 -4 mm2).

Conclusion

It was concluded that the migration of the endometrial cells to the extra-uterine environment (ovaries), higher proliferation rate and higher cell density of euescs in culture may be the responsible elements for the differences between euescs and eescs morphologies in endometriosis.

Keywords

Endometriosis, stromal cells, eutopic, ectopic, morphology