Basal characterization and in vitro differentiation of putative stem cells derived from the adult mouse ovary.

Lately, stem cell approaches have provided new information on reproductive organ function and additionally recommended novel treatment possibilities. The type(s) and differentiation potential of stem cells present in the mammalian ovary are largely unknown; while oogonial stem cells have been reported, we explored the possibility that multipotent stem cells may reside in the ovary and have wide differentiation potential. In this experimental study, homogenates of whole mouse ovaries were sorted using the stem cell surface markers stem cell antigen-1 and stage specific embryonic antigen-1/CD15. Viable double-positive cells 3-10 µm in diameter were evaluated immediately after sorting and after culture using differentiation conditions. Ovarian-derived stem cells were differentiated into the three main cell types: adipocytes, chondrocytes, or osteocytes. The subsequent culture was performed in media containing bone morphogenetic protein 4 (BMP-4) and/or retinoic acid (RA). RA, BMP-4 or the two agents in combination, consistently stimulated germ cell gene expression. RA treatment strongly stimulated germline gene expression and also the development of cells that were morphologically reminiscent of oocytes. The germ cell genes Dazl, Ddx4, Figla, Gdf-9, Nobox, Prdm9, and Sycp-1 were all detected at low levels. Remarkably, treatment with BMP-4 alone significantly increased protein expression of the granulosa cell product anti-Müllerian hormone (AMH). We have shown that an inclusive isolation protocol results in the consistent derivation of multipotent stem cells from the adult ovary; these cells can be differentiated towards the germ cell fate (RA alone), somatic ovarian cell fate as indicated by AMH production (BMP-4 alone), or classical mesenchymal cell types. Taken together, these data suggest the presence of multipotent mesenchymal stem cells in the murine ovary.

Etanercept protects ovarian reserve against ischemia/reperfusion injury in a rat model.

INTRODUCTION: Etanercept has been widely used in autoimmune diseases for blocking tumor necrosis factor α (TNF-α), which is an inflammatory cytokine. The anti-apoptotic and anti-inflammatory effects of etanercept against ischemia/reperfusion (I/R) injury have been shown for several tissues in rat studies, but to the best of our knowledge, there are no reports on its protective effects following similar injury in ovarian tissue. The aim of this study was to investigate whether etanercept has beneficial effects on ovarian I/R injury, as well as on ovarian reserve. MATERIAL AND METHODS: Twenty-four rats were randomly divided into four groups (n = 6/group): sham (laparotomy only); sham + etanercept; I/R; and I/R + etanercept. Ischemia was induced for 3 h by twisting the ovary, and 24 h after detorsion the ovarian tissues were collected to evaluate histopathologic changes, glutathione (GSH), malondialdehyde (MDA), myeloperoxidase (MPO), and superoxide dismutase (SOD) concentrations for oxidative stress, 8-hydroxy-2'-deoxyguanosine (8-OHdG) for DNA damage, caspase-3 activity for apoptosis and ovarian follicle counts. To measure anti-Mullerian hormone (AMH), serum samples were drawn before and after surgery. RESULTS: Tissue GSH and SOD levels were significantly higher, while MDA and MPO levels were significantly lower in the I/R + etanercept group than in the I/R group (p < 0.05, p < 0.01, respectively). Tissue 8-OHdG and caspase-3 activity were significantly lower in the I/R+etanercept group than in the I/R group (p < 0.05, p < 0.01, respectively). Preoperative and postoperative AMH levels were compared and there was a significant reduction in the I/R and I/R + etanercept groups (p < 0.001, p < 0.001). The reduction of AMH in the I/R + etanercept group was significantly lower than in the I/R group. The primordial, preantral and small antral follicle numbers were also significantly higher in the I/R + etanercept group compared to the I/R group (p < 0.001, p < 0.001, p < 0.005, respectively). CONCLUSIONS: Etanercept attenuated inflammation and related oxidative stress and also helped to preserve ovarian reserve following ovarian I/R damage.

Lipid profile and plasma atherogenic index in postmenopausal osteoporosis.

OBJECTIVE: The goal of this study was to investigate the relationship between the lipid profile, plasma atherogenic index (PAI), and osteoporosis in postmenopausal women. METHODS: The data of age, duration of menopause, height, weight, lipid profile, bone mineral density (BMD) value, and history of oral contraceptive use of 407 postmenopausal women who had not been menstruating for at least 12 months, were between the ages 45 and 80, and presented at the obstetrics and gynecology polyclinic of Kartal Dr. Lutfi Kirdar Tr aining and Research Hospital were reviewed. The patients were divided into 2 groups according to the presence of osteoporosis, and the data compared. The level of significance was accepted as p<0.05. RESULTS: A total of 142 postmenopausal patients with osteoporosis were included in the study. The mean age was 61.7±6.9 years. In the control group, there were 263 postmenopausal women without osteoporosis, with a mean age of 58.3±4.5 years. There was no statistically significant difference with respect to triglyceride level; however, in the osteoporosis group, the level of total cholesterol and low-density lipoprotein (LDL) were lower, and the level of high-density lipoprotein (HDL) was higher (p=0.762, p=0.002, p=0.01, p<0.001, respectively). CONCLUSION: A high level of HDL, and low LDL and PAI values, which are important for the prevention of cardiovascular disease, were found to be negative factors for BMD.

Medical Therapies for Endometriosis Differentially Inhibit Stem Cell Recruitment.

OBJECTIVE: To determine the effect of the 3 well-known endometriosis treatments on stem cell recruitment to endometriotic lesions. STUDY DESIGN: C57BL/6 mice (aged 8 weeks, n = 20) underwent bone marrow transplant following submyeloablation with 5-fluorouracil using 20 × 106 bone marrow stem cells from green fluorescent protein (GFP) mice. Two weeks after transplantation, experimental endometriosis was created in mice by suturing segments of the uterine horn into the peritoneal cavity. Mice were then randomized to receive treatment with medroxyprogesterone acetate (MPA), leuprolide acetate (Gonadotrophin-Releasing Hormone Analogue [GnRHa]), letrozole, or vehicle control (dimethyl sulfoxide). After 3 weeks of treatment, the mice were killed and the endometriosis lesions evaluated. RESULTS: All 3 treatments resulted in a significant reduction in lesion volume and weight. Estrogen deprivation using GnRHa or letrozole resulted in greater lesion regression than the progestin MPA. The GFP+/CD45- bone marrow-derived stem cells (BMDSCs) engrafted the lesions of endometriosis. Estrogen deprivation using GnRHa or letrozole significantly reduced BMDSC engraftment in the endometriosis lesions. MPA failed to significantly reduce stem cell number in endometriosis. CONCLUSION: The superiority of estrogen deprivation over progestin therapy in depriving the lesions of stem cells may have implications for the long-term treatment of endometriosis. Reduced stem cell engraftment is likely to result in long-term regression of the lesions, whereas progestins may only prevent their growth acutely.

Bone Marrow Stem Cell Chemotactic Activity Is Induced by Elevated CXCl12 in Endometriosis.

Endometriosis is an inflammatory gynecological disorder caused by the growth of endometrial tissue outside the uterus. Endometriosis produces chemokines, including CXCL12, that attract bone marrow cells to the lesions. In this study, we describe the expression, localization, and chemotactic activity of CXCL12 in endometriotic lesions. Biopsies were collected both from women with endometriosis undergoing laparoscopy and control endometrium from women without endometriosis. Expression of CXCl12 and CXCR4 messenger RNA was increased approximately 4- and 6-fold, respectively, in endometriosis compared to eutopic endometrium. Immunohistochemistry of lesions revealed that CXCR4 was expressed in the stroma and epithelium in both endometriosis and control eutopic endometrium. The level of CXCR4 protein expression was significantly higher in all cellular compartments of the endometriotic lesions compared to control endometrium. CXCL12 protein expression was also higher in endometriotic lesions and was greatest in the epithelial compartment. CXCL12 was increased more in the condition media of cultured endometriosis than in controls as measured by enzyme-linked immunosorbent assay. Transwell chamber migration was used to demonstrate 2-fold increased chemoattraction of mouse bone marrow stem cells toward CXCL12 in the endometriotic-conditioned medium compared with eutopic endometrium. Our results indicate that a preferential recruitment of stem cells to endometriosis can explain how endometriosis outcompetes eutopic endometrium in recruiting the limited supply of circulating stem cells. The CXCL12/CXCR4 signaling axis is a potential target for the treatment of endometriosis and its associated disorders.

Serum microRNAs as diagnostic markers of endometriosis: a comprehensive array-based analysis.

OBJECTIVE: To investigate serum microRNAs (miRNAs) in women with endometriosis. DESIGN: Case-control study. SETTING: University hospital. PATIENT(S): Women with (n = 24) and without (n = 24) endometriosis. INTERVENTION(S): Serum samples were obtained from surgically diagnosed subjects. MAIN OUTCOME MEASURE(S): miRNA from women with without endometriosis were used for microarray profiling and confirmed by means of quantitative real-time polymerase chain reaction (qRT-PCR). Receiver operating characteristic (ROC) analysis was performed on differentially expressed miRNAs. RESULT(S): miR-3613-5p, miR-6755-3p were down-regulated and miR-125b-5p, miR-150-5p, miR-342-3p, miR-143-3p, miR-145-5p, miR-500a-3p, miR-451a, miR-18a-5p were up-regulated more than 10-fold in the microarray. These results were confirmed with the use of qRT-PCR. Among the differentially expressed miRNAs, miR-125b-5p expression levels had the highest area under the ROC curve (AUC). The maximum AUC score of 1.000 was achieved when combining miR-125b-5p, miR-451a, and miR-3613-5p with the use of a logistic regression model. CONCLUSION(S): We identified several miRNAs in serum that distinguished subjects with endometriosis from those without. miR-125b-5p had the greatest potential as a single diagnostic biomarker. A combination of that miRNA with miR-451a and miR-3613-5p further improved diagnostic performance.

Is adjuvant radiotherapy necessary for FIGO stage 1a grade 2 endometrial endometrioid adenocarcinoma?

OBJECTIVE: The impact of adjuvant radiotherapy on the rates of survival and local recurrence was analyzed in patients diagnosed with International Federation of Gynecology and Obstetrics (FIGO) stage 1a grade 2 endometrial endometrioid adenocarcinoma. MATERIAL AND METHODS: Medical records of 82 patients diagnosed and treated for FIGO stage 1a grade 2 endometrial endometrioid adenocarcinoma were reviewed retrospectively. A group of 59 patients who received postoperative radiotherapy was compared with a control group of 23 subjects treated without adjuvant radiotherapy; the duration of survival as well as the local recurrence and metastasis rates were evaluated in both groups. RESULTS: The analysis of patient data has revealed the rate of local recurrence as 4.3% vs. 1.7% (p=0.485), the rate of distant metastasis as 4.3% vs. 6.9% (p=1.000), and the mean survival time as 83.6±38.7 vs. 81.5±37.5 months (p=0.828) in the adjuvant radiotherapy and control groups, respectively. CONCLUSION: In the presented study, adjuvant radiotherapy failed to improve the overall survival of the patients in the low-risk group (stage 1a grade 2). With the addition of the significant risk of radiation toxicity, it is highly probable that these patients will not benefit from postoperative radiotherapy. Close observation should be performed following the primary surgery in this patient group. Nevertheless, it should also be considered that adjuvant radiotherapy is a very effective treatment modality for the recovery of patients with vaginal relapse.