Volume 20, Issue 1 (2017)
mjms 2017, 20(1): 29-42 |
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Rahimipour M, Salehnia M, Jafarabadi M. Alteration of Genes Expression Related to Implantation in Human Endometrial Stromal Cells Cultured in the Presence of Ovarian Hormones. mjms 2017; 20 (1) :29-42
URL: http://mjms.modares.ac.ir/article-30-511-en.html
URL: http://mjms.modares.ac.ir/article-30-511-en.html
1- Department of Anatomy, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
2- Reproductive Health Research Center, Tehran University of Medical Sciences, Tehran, Iran
2- Reproductive Health Research Center, Tehran University of Medical Sciences, Tehran, Iran
Abstract: (8272 Views)
Objective: This experimental study evaluatedthe effect of ovarian hormones on the expression of genes related to implantation in human endometrial stromal cells in vitro.
Methods: After confirmation ofnormal endometrial tissue collected from hysteroscopy according to hematoxylin and eosin staining, we isolated the endometrial cells with collagenase type 1. The cells were passed through 100 and 40 µm filters. We cultured the collected stromal cells in DMEM/F-12 medium to the fourth passage. Passage-4 cells were examined for 7 days in two groups - control (without hormones) and experimental groups with different concentrations of estrogen (E2; 0.3, 0.7 and 1 nmol/L) and a constant concentration of progesterone (P4; 63.5 nmol/L). After culture, the cell viability, proliferation and the expression of genes related to implantation, αv and β3 integrins, leukemia inhibitory factor receptor (LIFR), and interleukin-1 receptor (IL-1R) were assessed by staining with propidium iodide and acridine orange, the MTT assay, and real time RT-PCR.The data were analyzed using SPSS software.
Results: There were 99.9% viable cells in all groups. Cell proliferation increased significantly in the 0.3 nmol/L treated group compared with the control group and the group treated with 0.7nmol/L (P≤0.04). However, the group treated with 1 nmol/L did not significantly change compared to the other groups. In contrast to IL1-R and LIFR, expressions of the αv and β3 integrins significantly increased in the 0.3 nmol/L treated group compared to the control (P≤0.02).
Conclusion: The combined use of estrogen and progesterone in an endometrial stromal cell culture had an effect on the proliferation and expression of some genes related to implantation.
Methods: After confirmation ofnormal endometrial tissue collected from hysteroscopy according to hematoxylin and eosin staining, we isolated the endometrial cells with collagenase type 1. The cells were passed through 100 and 40 µm filters. We cultured the collected stromal cells in DMEM/F-12 medium to the fourth passage. Passage-4 cells were examined for 7 days in two groups - control (without hormones) and experimental groups with different concentrations of estrogen (E2; 0.3, 0.7 and 1 nmol/L) and a constant concentration of progesterone (P4; 63.5 nmol/L). After culture, the cell viability, proliferation and the expression of genes related to implantation, αv and β3 integrins, leukemia inhibitory factor receptor (LIFR), and interleukin-1 receptor (IL-1R) were assessed by staining with propidium iodide and acridine orange, the MTT assay, and real time RT-PCR.The data were analyzed using SPSS software.
Results: There were 99.9% viable cells in all groups. Cell proliferation increased significantly in the 0.3 nmol/L treated group compared with the control group and the group treated with 0.7nmol/L (P≤0.04). However, the group treated with 1 nmol/L did not significantly change compared to the other groups. In contrast to IL1-R and LIFR, expressions of the αv and β3 integrins significantly increased in the 0.3 nmol/L treated group compared to the control (P≤0.02).
Conclusion: The combined use of estrogen and progesterone in an endometrial stromal cell culture had an effect on the proliferation and expression of some genes related to implantation.
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