Leukemia inhibitory factor increases the proliferation of human endometrial stromal cells and expression of genes related to pluripotency

Background: concerning the low population of human endometrial mesenchymal cells within the tissue and their potential application in the clinic and tissue engineering, some researches have been focused on their in vitro expansion

Objective: the aim of this study was to evaluate the effect of leukemia inhibitory factor [LIF] as a proliferative factor on the expansion and proliferation of human endometrial stromal cells

Materials and Methods: in this experimental study, the isolated and cultured human endometrial stromal cells from women at ovulatory phase aged 20-35 years, after fourth passage were divided into control and LIF-treated groups. In the experimental group, the endometrial cells were treated by 10 ng/ml LIF in culture media and the cultured cells without adding LIF considered as control group. Both groups were evaluated and compared for proliferation rate using MTT assay, for CD90 marker by flow cytometric analysis and for the expression of Oct4, Nanog, PCNA and LIFr genes using real-time RT-PCR

Results: the proliferation rate of control and LIF-treated groups were 1.17+/-0.17 and 1.61+/-0.06 respectively and there was a significant increase in endometrial stromal cell proliferation following in vitro treatment by LIF compared to control group [p=0.049]. The rate of CD90 positive cells was significantly increased in LIFtreatedgroup [98.96+/-0.37%] compared to control group [94.26+/-0.08%] [p=0.0498]. Also, the expression ratio of all studied genes was significantly increased in the LIFtreated group compared to control group [p=0.0479]

Conclusion: the present study showed that LIF has a great impact on proliferation, survival, and maintenance of pluripotency of human endometrial stromal cells and it could be applicable in cell therapies

Characteristics of human endometrial stem cells in tissue and isolated cultured cells: an immunohistochemical aspect

Background: The aim of this study was to investigate the percentage of the stem cells population in human endometrial tissue sections and cultured cells at fourth passage

Methods: Human endometrial specimens were divided into two parts, one part for morphological studies and the other part for in vitro culture. Full thickness of human normal endometrial sections and cultured endometrial cells at fourth passage were analyzed via immunohistochemistry for CD146 and some stemness markers such as Oct4, Nanog, SoX[2], and Klf4 and the expression of typical mesenchymal stem cell markers CD90, CD105

Results: 11.88 +/- 1.29% of human endometrial cells within tissue sections expressed CD146 marker vs. 28 +/- 2.3% of cultured cells, CD90 and CD105 were expressed by functionalis stroma [85 +/- 2.4 and 89 +/- 3.2%] than basalis stroma [16 +/- 1.4 and 17 +/- 1.9%], respectively [P<0.05]. Oct4 and Nanog-expressing cells comprise 1.43 +/- 0.08 and 0.54 +/- 0.01% of endometrial stromal cells in endometrial sections vs. 12 +/- 3.1% and 8 +/- 2.9% of cultured cells, respectively. They reside near the glands in the basal layer of endometrium. SoX[2] and Klf4 were not commonly expressed in tissue samples and cultured cells. CD9 and EpCAM were expressed by epithelial cells of the endometrium, rather than by stroma or perivascular cells

Conclusion: The human endometrial stem cells and pluripotency markers may be localized more in basalis layer of endometrium. The immunostaining observations of endometrial cells at fourth passage were correlated with the immunohistochemistry data

effect of stem cell factor on proliferation of human endometrial CD146+ cells

Background: Stem cell factor [SCF] is a transcriptional factor which plays crucial roles in normal proliferation, differentiation and survival in a range of stem cells

Objective: The aim of the present study was to examine the proliferation effect of different concentrations of SCF on expansion of human endometrial CD146+ cells

Materials and Methods: In this experimental study, total populations of isolated human endometrial suspensions after fourth passage were isolated by magnetic activated cell sorting [MACS] into CD146+ cells. Human endometrial CD146+ cells were karyotyped and tested for the effect of SCF on proliferation of CD146+ cells, then different concentrations of 0, 12.5, 25, 50 and 100 ng/ml was carried out and mitogens-stimulated endometrial CD146+ cells proliferation was assessed by MTT assay

Results: Chromosomal analysis showed a normal metaphase spread and 46XX karyotype. The proliferation rate of endometrial CD146P + P cells in the presence of 0, 12.5, 25, 50 and 100 ng/ml SCF were 0.945 +/- 0.094, 0.962 +/- 0.151, 0.988 +/- 0.028, 1.679 +/- 0.012 and 1.129 +/- 0.145 respectively. There was a significant increase in stem/ stromal cell proliferation following in vitro treatment by 50 ng/ml than other concentrations of SCF [p=0.01]

Conclusion: The present study suggests that SCF could have effect on the proliferation and cell survival of human endometrial CD146P+P cells and it has important implications for medical sciences and cell therapies

Ovarian stimulation by exogenous gonadotropin decreases the implantation rate and expression of mouse blastocysts integrins

Integrins are heterodimeric glycoprotein receptors that regulate the interaction of cells with extracellular matrix and may have a critical role in implantation. The aim of this study was to investigate the effect of ovulation induction on the expression of Alpha4, Alphav, Beta1, and Beta3 integrins in mouse blastocyst at the time of implantation. The ovarian stimulated and non-stimulated pregnant mice were sacrificed on the morning of 5[th] day of pregnancy. The blastocysts were collected, and the expression of Alphav, Alpha4, Beta1, and Beta3 integrins was examined using real-time RT-PCR and immunocytochemical techniques, then their ovarian hormones were analyzed at the same time. The implantation sites in uterine horns of other pregnant mice in both groups were determined under a stereomicroscope on the 7[th] day of pregnancy. The results showed that the expression of Alphav, Beta1, and Beta3 integrins in both mRNA and protein levels was significantly lower in the ovarian stimulated group than the control group, and the maximum ratio of expression was belonged to Beta1 molecule [P>0.05]. The implantation rate in superovulated mice was significantly lower than control mice. It was suggested that ovulation induction decreased the expression of Alphav, Beta1, and Beta3 integrins of mouse blastocysts

Assessment of alpha4, alphav, beta1 and beta3 integrins expression throughout the implantation window phase in endometrium of a mouse model of polycystic ovarian syndromes

Endometrial integrin expression changes might be a reason for implantation failure in polycystic ovarian syndromes [PCOS]. Assessment of integrin genes and proteins expression upon endometrium in the PCOS experimental mouse model was the main goal of this study. 30 NMRI female mice were equally divided into control, experimental [PCOS; received estradiol valerate [40 mg/kg]] and sham group [received; olive oil]. After 8 weeks, each group was hyper stimulated by 7 IU PMSG and then, after 48hrs, 7 IU HCG was injected. Vaginal plaque was checked. After 5 days, Progesterone and estradiol levels and endometrial tissues were investigated to evaluate of alpha4, alphav, beta1 and beta3 integrins gene and protein by qPCR method and immunohistochemistry, respectively. Tissue samples were assessed and showed that level of progesterone was significantly decreased in PCOS group. Results of molecular part in the amount of alphav, beta3, beta1 and alpha4 gene expressions showed a great difference in beta3 and alphav genes expressions between experimental groups, alphav, beta3, alpha4 and beta1 proteins in the endometrial stroma in the control group were expressed, but they were not detected in PCOS group. According to the results, integrins had different expression patterns in different areas of the endometrium; such as epithelial and stromal. It seems that in PCOS, this pattern has changed and the results might have a great influence on implantation failure. Therefore, this study suggests that a great attention to this problem may be essential in patients who are involved

Expression of alpha4, alphav, beta1 and beta3 integrins during the implantation window on blastocyst of a mouse model of polycystic ovarian syndromes

It has been hypothesized that blastocyst integrin expression changes can affect the spontaneous miscarriage in polycystic ovarian syndromes [PCOS]. In this study, the profile of integrin genes and proteins was investigated on blastocyst of the PCOS experimental mouse model. 30 NMRI female mice were equally divided into 3 groups: control, experimental [PCOS that was injected estradiol valerate [40 mg/kg]]. After 8 weeks, each group was hyper stimulated by PMSG and HCG. Vaginal plaque was checked, and mice were investigated 5 days after the test. Progesterone and estradiol levels were determined; alpha4, alpha v, beta 1 and beta 3 integrin genes and protein of blastocysts were examined by real time PCR method and immunohistochemistry, respectively. Estradiol level was significantly increased [p