The Effect of Different Doses of Clomiphen Citrate on Morphology and Proliferation of Human Endometrial Stromal Cells in In-Vitro Culture

Abstract

BACKGROUND AND OBJECTIVE: Clomiphene citrate is a drug which used for superovulation in infertile women. Despite increasing oocyte number, its pregnancy rate is low. Endometrial stromal cells have pivotal role in implantation events. The aim of present investigation was to determine the effect of different clomiphene citrate doses on morphology and proliferation of human endometrial stromal cell in in-vitro culture. METHODS: In this experimental study, human endometrial biopsies (n=7) chopped in sterile condition and digested by enzymatic methods (collagenase). Stromal cell harvested after using cell strainer and centrifugation. Their viability and cell number were determined with Trypan blue staining. Stromal cells divided into four groups: control and cases: (1, 5 and 10 µM clomiphene). Control group received DMEM/F12 media supplemental with FBS (5%) and antibiotic and cases groups received same media with one dose of clomiphene citrate. Duration of study was two weeks and cells were photographed at beginning and the end of study. Finally cell collected enzymaticly by tripsin (0.25%) and their viability and cell number were determined. FINDINGS: Morphological changes resembled in control and 1 µM groups and consist of oval cell transformation into large polyclonal fibroblast. 5 and 10µM doses of clomiphene citrate inhibit normal proliferation and morphological changes. There are significant differences between 5 and 10µM with control and 1µM groups (p<0.001).CONCLUSION: Clomiphene citrate showed dose dependant effect on endometrial stromal cell and higher doses inhibit proliferation and morphological changes of these cells.

Keywords

• human endometrium
• stromal cell
• cell culture
• clomiphene citrate