RESUMEN
This study aims to evaluate the effect of melatonin supplementation on the outcomes of in vitro fertilization (IVF) and mitochondrial adenosine triphosphate production (MT-ATP6) gene expression in Iranian infertile couples. A single-blind nonrandomized controlled trial was conducted, recruiting 90 infertile couples who underwent IVF at an infertility center in Tehran, Iran. Patients who were assigned to the intervention group received melatonin as a supplementation to the standard controlled ovarian stimulation (COS). The control group received a COS protocol only. Primary outcome was the mRNA level of the MT-ATP6 gene in cumulus cells of ovarian follicles. Secondary outcomes were the mean number of mature oocytes retrieved, the embryo quality, and biochemical and clinical pregnancy rates. The mRNA level of the MT-ATP6 gene in cumulus cells between intervention and control groups was not statistically different (0.931 vs.1; P Ë 0.05). The mean number of poor-quality embryos was significantly lower in the intervention group than that in the control group (0.27 vs. 0.80; P = 0.028). The biochemical and clinical pregnancy rates were higher in the intervention group (24% vs. 14%, P = 0.089, and 14% vs. 7%, P = 0.302, respectively); however, the difference was not significant. Melatonin supplementation did not increase the odds of clinical pregnancy and the number of mature oocytes retrieved, but significantly reduced the number of low-quality embryos. More extensive studies focusing on the level of MT-ATP6 gene expression in the oocyte or blastomere cells may further elucidate the effect of supplementation with melatonin in infertile couples who have poor clinical outcomes. Trial registration: Current Controlled Trials: IRCT2015042912307N4.
Asunto(s)
Fertilización In Vitro/tendencias , Infertilidad/metabolismo , Infertilidad/terapia , Melatonina/administración & dosificación , ATPasas de Translocación de Protón Mitocondriales/biosíntesis , Índice de Embarazo/tendencias , Administración Oral , Adulto , Antioxidantes/administración & dosificación , Células del Cúmulo/efectos de los fármacos , Células del Cúmulo/metabolismo , Femenino , Fertilización In Vitro/métodos , Expresión Génica , Humanos , Infertilidad/epidemiología , Irán/epidemiología , Masculino , ATPasas de Translocación de Protón Mitocondriales/genética , Embarazo , Método Simple Ciego , Resultado del TratamientoRESUMEN
Recently, due to tremendous progress in prognosis, diagnosis, and treatment of different kinds of malignancies, demands on fertility preservation were raised significantly in developed countries. Fertility failure is one of the most detrimental consequences of radio/cytotoxic treatment procedures in women who could overcome their cancer disease. For women who are involved in cancer diseases, there are multiple options regarding their fertility preservation and those could be selected according to patient's age, the risk of ovarian involvement, the available time and the type of cancer with different levels of advantages and disadvantages. Although there are multiple options, but embryo cryopreservation and ovarian tissue cryopreservation are the most reliable methods for permature and post-mature puberty, respectively. In addition, other approaches like artificial ovary, isolation and cryopreservation of follicles and mature and immature oocyte preservation are under investigations and the success rate of oocyte vitrification is increasing. Therefore, the techniques have the potential to be used in clinic in near future. The presence of comprehensive consultation, before the onset of any kind of cancer treatment procedures, is an indispensable issue which would help patients to make up their mind in choosing the immediate and the best available fertility preservation option.
RESUMEN
One of the most important concerns cancer survivors face is fertility. Current treatment modalities often result in damage to the reproductive system. Different options have been proposed to preserve the fertility of affected women, and many attempts have been made to improve their chance of childbearing after therapy. Cryopreservation of ovarian tissue and follicles before the onset of cancer treatment and then either transplantation of ovarian tissue or culture of ovarian tissue and individual follicles in vitro is a commonly cited approach. Extensive research is being done to design an optimal condition for the culture of ovarian follicles. Improving follicle culture systems by understanding their actual growth needs might be a crucial step toward fertility preservation in cancer patients. This review article will try to provide a summary of the role of different factors and conditions on growth of human and bovine preantral follicles in vitro.
Asunto(s)
Criopreservación/métodos , Preservación de la Fertilidad/métodos , Neoplasias , Folículo Ovárico/citología , Folículo Ovárico/crecimiento & desarrollo , Animales , Bovinos , Femenino , Humanos , Técnicas In Vitro , Oocitos/citología , Oocitos/crecimiento & desarrolloRESUMEN
Follicle culture provides a condition which can help investigators to evaluate various aspects of ovarian follicle growth and development and impact of different components and supplementations as well as presumably application of follicle culture approach in fertility preservation procedures. Mesenchymal Stem Cells (MSCs), particularly those isolated from menstrual blood has the potential to be used as a tool for improvement of fertility. In the current study, a 3D co-culture system with mice preantral follicles and human Menstrual Blood Mesenchymal Stem Cells (MenSCs) using either collagen or alginate beads was designed to investigate whether this system allows better preantral follicles growth and development. Results showed that MenSCs increase the indices of follicular growth including survival rate, diameter, and antrum formation as well as the rate of in vitro maturation (IVM) in both collagen and alginates beads. Although statistically not significant, alginate was found to be superior in terms of supporting survival rate and antrum formation. Hormone assay demonstrated that the amount of secreted 17 ß-estradiol and progesterone in both 3D systems increased dramatically after 12â¯days, with the highest levels in system employing MenSCs. Data also demonstrated that relative expression of studied genes increased for Bmp15 and Gdf9 and decreased for Mater when follicles were cultured in the presence of MenSCs. Collectively, results of the present study showed that MenSCs could improve indices of follicular growth and maturation in vitro. Further studies are needed before a clinical application of MenSCs-induced IVM is considered.
Asunto(s)
Células Madre Adultas/citología , Menstruación , Células Madre Mesenquimatosas/citología , Oogénesis , Folículo Ovárico/citología , Andamios del Tejido , Adulto , Células Madre Adultas/metabolismo , Alginatos/química , Animales , Biomarcadores/metabolismo , Células de la Médula Ósea/citología , Diferenciación Celular , Supervivencia Celular , Células Cultivadas , Técnicas de Cocultivo , Colágeno/química , Femenino , Preservación de la Fertilidad , Ácido Glucurónico/química , Ácidos Hexurónicos/química , Humanos , Técnicas de Maduración In Vitro de los Oocitos , Células Madre Mesenquimatosas/metabolismo , Ratones , Microesferas , Folículo Ovárico/metabolismo , Técnicas de Cultivo de Tejidos , Ingeniería de Tejidos , Andamios del Tejido/químicaRESUMEN
The recent discovery of ovarian stem cells in postnatal mammalian ovaries, also referred to as putative stem cells (PSCs), and their roles in mammalian fertility has challenged the long-existing theory that women are endowed with a certain number of germ cells. The rare amount of PSCs is the major limitation for utilizing them through different applications. Therefore, this study was conducted in six phases to find a way to increase the number of Fragilis- and mouse vasa homolog (MVH)-positive sorted cells from 14-day-old NMRI strain mice. Results showed that there is a population of Fragilis- and MVH-positive cells with pluripotent stem cell characteristics, which can be isolated and expanded for months in vitro. PSCs increase their proliferation capacity under the influence of some mitogenic agents, and our results showed that different doses of stem cell factor (SCF) induce PSC proliferation with the maximum increase observed at 50 ng/mL. SCF was also able to increase the number of Fragilis- and MVH-positive cells after sorting by magnetic-activated cell sorting and enhance colony formation efficiency in sorted cells. Differentiation capacity assay indicated that there is a basic level of spontaneous differentiation toward oocyte-like cells during 3 days of culture. However, relative gene expression was significantly higher in the follicle-stimulating hormone-treated groups, especially in the Fragilis- sorted PSCs. We suggest that higher number of PSCs provides us either a greater source of energy that can be injected into energy-impaired oocytes in women with a history of repeat IVF failure or a good source for research.
Asunto(s)
Diferenciación Celular , Proliferación Celular , Separación Celular/métodos , Oocitos/citología , Ovario/citología , Células Madre Pluripotentes/citología , Animales , Técnicas de Cultivo de Célula , Células Cultivadas , Femenino , Hormona Folículo Estimulante/farmacología , Ratones , Oocitos/efectos de los fármacos , Ovario/efectos de los fármacos , Células Madre Pluripotentes/efectos de los fármacosRESUMEN
Placenta harbors a plentiful source of various cells with stem cells or stem-like cell properties, which can be used in therapeutic procedures and research. Mesenchymal stem cells (MSCs) have attracted much attention due to their specific differentiation potential and tolerogenic properties. MSCs have been isolated from different parts of placenta; however, in this study, we isolated MSCs from amnion and chorion membrane, as well as umbilical cord (Wharton's jelly [WJ]) and compared their capacity regarding differentiation toward female germ cells under influence of 10 ng/mL BMP4. All placenta samples were collected from delivering mothers by normal cesarean section and cells were isolated by different methods. Results showed that all isolated cells were mostly positive for the MSC markers CD73, CD166, and CD105, and minimally reacted with CD34 and CD45 (hematopoietic markers). After differentiation induction using third passage cultured cells, immunocytochemistry staining showed that cells were positive for germline cell-related genes Ssea4, Oct4, and Ddx4, and oocyte-related gene Gdf9. RT-qPCR results indicated that human chorion MSCs (hCMSCs) had a greater potential to be differentiated into female germline cells. Moreover, the results of this study indicate that human umbilical cord MSCs originated from either male or female umbilical cord have the same differentiation potential into female germline cells. We recommend that for presumptive application of MSCs for infertility treatment and research, hUMSCs are best candidates due to their higher differentiation potential, ease of proliferation and expansion, and low immunogenicity.
Asunto(s)
Amnios/citología , Diferenciación Celular , Corion/citología , Células Germinativas/citología , Células Madre Mesenquimatosas/citología , Placenta/citología , Cordón Umbilical/citología , Células Cultivadas , Femenino , Humanos , EmbarazoRESUMEN
Cryopreservation of spermatogonial stem cells is considered as a useful procedure for preserving fertility in children with testis cancer. SSCs were isolated from testes mice, and then antioxidant was added to the freezing medium. The Bax expression level in antioxidant groups was significantly (P ≤ 0.05) lower than the control group and a significant rise of Bcl2 expression was detected in the antioxidant groups. ROS production with antioxidant was significantly lower compared with the control group. Cryopreservation with the addition of the antioxidants can help increase the number of SSCs and improve the quality and viability of these cells after cryopreservation.
Asunto(s)
Células Madre Germinales Adultas/metabolismo , Antioxidantes/farmacología , Catalasa/farmacología , Criopreservación , Estrés Oxidativo/efectos de los fármacos , alfa-Tocoferol/farmacología , Células Madre Germinales Adultas/citología , Animales , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Congelación , Masculino , RatonesRESUMEN
An increasing body of evidence has confirmed existence and function of ovarian stem cells (OSCs). In this study, a novel approach on differentiation of OSCs into oocyte-like cells (OLCs) has been addressed. Recently, different methods have been recruited to isolate and describe aspects of OSCs, but newer and more convenient strategies in isolation are still growing. Herein, a morphology-based method was used to isolate OSCs. Cell suspension of mouse neonatal ovaries was cultured and formed colonies were harvested mechanically and cultivated on mouse embryonic fibroblasts. For differentiation induction, colonies transferred on inactive granulosa cells. Results showed that cells in colonies were positive for alkaline phosphatase activity and reverse transcription-polymerase chain reaction (RT-PCR) confirmed the pluripotency characteristics of cells. Immunofluorescence revealed a positive signal for OCT4, DAZL, MVH, and SSEA1 in colonies as well. Results of RT-PCR and immunofluorescence confirmed that some OLCs were generated within the germ stem cell (GSCs) colonies. The applicability of morphological selection for isolation of GSCs was verified. This method is easier and more economic than other techniques. Our results demonstrate that granulosa cells were effective in inducing the differentiation of OSCs into OLCs through direct cell-to-cell contacts.
Asunto(s)
Técnicas de Cultivo de Célula/métodos , Diferenciación Celular , Células de la Granulosa/citología , Oocitos/citología , Ovario/citología , Células Madre/citología , Animales , Células Cultivadas , Técnicas de Cocultivo , Femenino , RatonesRESUMEN
OBJECTIVES: Several researchers have reported the relationship between infertility in male and varicocele for so many years but the implication of varicocele in female patients is remains elusive. Here, we aim to examine the effects of unilateral varicose ovarian vein on antioxidant capacity and oocyte quality of rat ovary after the experimental creation of varicocele in female rats. MATERIALS AND METHODS: In this study, thirty adult female albino rats were divided into three equal groups: Group 1 as the control group has 10 rats, Group 2 as the sham group has 10 rats and they underwent a sham operation and finally Group 3 has the varicocele group has 10 rats. Antioxidant assays for superoxide dismutase, glutathione peroxidase and catalase were performed using specific assay kits and gene expression for Bax, Bmp-15, Hsp-27 and Gdf-9 was done via real time PCR. RESULTS: The adverse effects of the experimentally induced varicocele were reported and recorded on the left ovary compared to the right sided ovary (no varicocele induction) in the varicocele group. Real time PCR data shows that the expression of Gdf-9, Hsp-27 and Bmp-15 genes were all significantly reduced at p≤ 0.05. CONCLUSION: The results of this study show that reduced gene expression of Bmp-15, Gdf-9 and Hsp-27, increased gene expression of bax and an imbalance between pro-oxidant/antioxidant ratio are few of the several mechanisms by which varicocele may lead to infertility in female.
RESUMEN
Although existence of ovarian stem cells (OSCs) in mammalian postnatal ovary is still under controversy, however, it has been almost accepted that OSCs are contributing actively to folliculogenesis and neo-oogenesis. Recently, various methods with different efficacies have been employed for OSCs isolation from ovarian tissue, which these methods could be chosen depends on aim of isolation and accessible equipments and materials in lab. Although isolated OSCs from different methods have various traits and characterizations, which might become from their different nature and origin, however these stem cells are promising source for woman infertility treatment or source of energy for women with a history of repeat IVF failure in near future. This review has brought together and summarized currently used protocols for isolation and propagation of OSCs in vitro.