An Overview of The Available Methods for Morphological Scoring of Pre-Implantation Embryos in In Vitro Fertilization

Assessment of embryo quality in order to choose the embryos that most likely result in pregnancy is the critical goal in assisted reproductive technologies (ART). The current trend in human in vitro fertilization/embryo transfer (IVF/ET) protocols is to decrease the rate of multiple pregnancies after multiple embryo transfer with maintaining the pregnancy rate at admissible levels (according to laboratory standards). Assessment of morphological feathers as a reliable non-invasive method that provides valuable information in prediction of IVF/intra cytoplasmic sperm injection (ICSI) outcome has been frequently proposed in recent years. This article describes the current status of morphological embryo evaluation at different pre-implantation stages

Downregulation Of Extracellular Matrix And Cell Adhesion Molecules In Cumulus Cells Of Infertile Polycystic Ovary Syndrome Women With And Without Insulin Resistance

Objective The extracellular matrix (ECM) of the cumulus oocyte complex (COC) is composed of several molecules that have different roles during follicle development. This study aims to explore gene expression profiles for ECM and cell adhesion molecules in the cumulus cells of polycystic ovary syndrome (PCOS) patients based on their insulin sensitivity following controlled ovarian stimulation (COS). Materials And Methods In this prospective case-control study enrolled 23 women less than 36 years of age who participated in an intracytoplasmic sperm injection (ICSI) program. Patients were subdivided into 3 groups: control (n=8, fertile women with male infertility history), insulin resistant (IR) PCOS (n=7), and insulin sensitive (IS) PCOS (n=8). We compared 84 ECM component and adhesion molecule gene expressions by quantitative real-time polymerase chain reaction array (qPCR-array) among the groups. Results We noted that 21 of the 84 studied genes differentially expressed among the groups, from which 18 of these genes downregulated. Overall, comparison of PCOS cases with controls showed downregulation of extracellular matrix protein 1 (ECM1); catenin (cadherin-associated protein), alpha 1 (CTNNA1); integrin, alpha 5 (ITGA5); laminin, alpha 3 (LAMA3); laminin, beta 1 (LAMB1); fibronectin 1 (FN1); and integrin, alpha 7 (ITGA7). In the IS group, there was upregulation of ADAM metallopeptidase with thrombospondin type 1 motif, 8 (ADAMTS8) and neural cell adhesion molecule 1 (NCAM1) compared with the controls (P<0.05). Conclusion Downregulation of ECM and cell adhesion molecules seem to be related to PCOS. Gene expression profile alterations in cumulus cells from both the IS and IR groups of PCOS patients seems to be involved in the composition and regulation of ECM during the ovulation process. This study highlights the association of ECM gene alteration as a viewpoint for additional understanding of the etiology of PCOS

Mesenchymal Stem Cells: In Vitro Differentiation among Bone and Cartilage Cell Lineages

Received: 1/May/2007, Accepted: 5/Jul/2007The capacity of mesenchymal stem cells (MSCs) to differentiate amongskeletal cell lineages and to undergo extensive proliferation in vitro rendersthem an appropriate source for cell-replacement therapy to heal defects ofbone and cartilage tissue.It is argued that in bone and cartilage defects,MSCs would better be transplanted as fully-differentiated cells otherwise theymay produce non-specific cells in defect sites. This notion may emphasizethe importance of the studies considering in vitro bone and cartilagedifferentiation of MSCs. Indeed, the capacity of producing osteoblastic andchondrocytic cell lineage is among the earliest differentiation potentials ofMSCs, being reported at first isolation of the cells. Recent studies, howeverindicate that MSCs could differentiate into more cell lineages than expected.The present study provides evidence for, in vitro potential of MSCs todifferentiate into bone and cartilage cell lineages. As an introduction tothe differentiation, the characteristics of MSCs have been described andMSCs differentiation reviewed. The culture condition for bone and cartilagedifferentiation, molecular regulation of the differentiation, signaling pathwayinvolved during the differentiation, and the genes up-regulated upon boneand cartilage differentiation have also been described

The structure of Human Mesenchymal Stem Cells Differentiated into Cartilage in Micromass Culture System

Introduction: The aim of this study was to differentiate humanmesenchymal stem cells (hMSCs) into cartilage in a micromass culturesystem and study of their structure by light and electron microscopy.Material and Methods: Human bone marrow cells obtained from volunteerpatients were plated in 75-cm2 flasks and their MSCs were expandedthrough several sub-cultures. The passage 4 cells were used to establishmicromass culture system for chondrogenic differentiation. For this purpose,200,000 fibroblastic cells were placed in centrifuge tubes and pelleted at 250g for 5 minutes. About 0.5 ml chondrogenic induction medium was thenadded to the pellet and the culture incubated in 5% CO2 at 37°C for 21 days.Then, some pellets were utilized to evaluate chondrogenic differentiation byeither RT-PCR analysis of some cartilage marker molecules or specificstaining for detecting cartilage matrix, and other pellets were used for lightand electron microscopic study of differentiated tissue.Results: Primary culture of the bone marrow cells were initially composed ofthe spindle- and round shaped cells, from which the spindle cells remainedand expanded through several passages. At the end of differentiation period,RT-PCR analysis showed high production of collagen II and X and aggrecanmRNA inside the differentiated cells, and toluidine blue staining indicatedintermediate accumulation of the metachromatic matrix among the inducedcells. In general, light micrograph indicated a rather cellular state of thedifferentiated tissue in which the peripheral part had more metachromaticmatrix than central zone. More detailed study of the sections revealed thatinduced aggregates of the cells were composed externally of very thin layerof elongated cells reminiscent of perichondrium and internally a mass of ovalcells comprising the main part of the pellet. Ultra-thin sections showed thatthe cells in perichondrium-like layer were very similar to fibroblastic cells andthose located centrally had a set of well-developed organelles, characteristicof highly active cells. Some fat cells were seen in central zone.Conclusion: Cartilage tissue differentiated from MSCs in micromass culturesystem seemed to be structurally very similar to developing cartilage not toadult mature cartilage

Murine Adherent CD34+ Cell Population Expanded by Single Cell Cloning

Introduction: While human endothelial progenitor cells (EPCs) have been a subject of somehow extensive investigation, EPCs from adult mouse hematopoietic system were poorly studied. Present investigation is focused on FVB mouse endothelial progenitor cells in terms of their isolation, purification, and expansion. Materials and Methods: Mononuclear cells collected from murine peripheral blood were cultured in fibronectin coated plate for two weeks, at which point, the adherent cell population were lifted and analyzed in terms of some surface markers. Using FACS Vantage equipped with one-cell deposition unit, single CD34 positive cells were plated per well already containing medium optimized for single cell growth. Several clones were then emerged, expanded, and examined in terms of some surface markers. Furthermore, the cells were investigated regarding ability to uptake DiI-ac-LDL and form capillary network on matrigel surfaces. Results: Adherent population of mononuclear cells from mouse peripheral blood was appeared morphologically heterogeneous. About 5% of the adherent cells were CD34 positive. Having optimized their culture condition, several CD34 positive clones were expanded. The cells comprising the clones were DiI-ac-LDL+ and formed capillary-like tube when being seeded on matrigel surfaces. Conclusion: The primary culture of the mononuclear cells from murine peripheral blood contains a very limited number of cells positive for endothelial lineage markers. These cells (adherent CD34 positive) could be expanded by single cell cloning technique

Is Coasting Valuable in All Patients with Any Cause of Infertility?

Objectives: This study aimed to assess the influence of coasting duration on the number and quality of oocytes and fertilization rate in male factor infertile women and those with polycystic ovary syndrome (PCOS). Methods: In this prospective observational follow-up study, 114 patients undergoing coasting (53 women with male factor infertility and 61 women with PCOS) were evaluated at the Royan Institute Research Center, Iran, between 2010 and 2012. Results: The results were analyzed according to the coasting periods of 1–4 days. In normal females, the number of oocytes retrieved was significantly reduced after the second day (p = 0.004). In addition, a statistically significant drop was observed in the number of metaphase II oocytes and fertilization rate after the third day (p = 0.006 and p = 0.006, respectively). No significant differences were observed in the number and quality of oocytes retrieved and fertilization rate with regard to coasting days in PCOS patients. Conclusion: Coasting with duration of more than three days should be performed with caution in normal females who are at risk of developing ovarian hyperstimulation syndrome

Oxidative Stress Statues in Serum and Follicular Fluid of Women with Endometriosis

Objective: This study aimed to evaluate the levels of two oxidative stress (OS) markers including lipid peroxide (LPO) and total antioxidant capacity (TAC) in both serum and follicular fluid (FF) of women with endometriosis after puncture. Materials and Methods: In this cross-sectional study, a total number of sixty-three women younger than 40 years old with laparoscopy (gold standard for endometriosis diagnosis) indication underwent in vitro fertilization (IVF) program in the Royan Institute, Tehran, Iran from September 2013 to October 2014. About forty-three patients were diagnosed with endometriosis after laparoscopy. Blood and FF from the leading follicle in each stimulated ovary were obtained at the time of egg retrieval; samples were centrifuged and frozen until assessment. At the time of sample assessment, serum and FF samples were evaluated for the levels of LPO and TAC on spectrophotometery. Results: We observed that women with endometriosis had significantly higher LPO and lower TAC levels in the serum and FF as compared with the control group (P<0.05). Conclusion: It has observed that FF of women with endometriosis, regardless of disease stage, increases the proliferation power of endometrial cells in vitro, we presume that inflammatory reactions-induced OS in ovary may be responsible for proliferation induction ability in FF obtained from women with endometriosis

Effects of Saffron (Crocus sativus L.) Aqueous Extract on In vitro Maturation, Fertilization and Embryo Development of Mouse Oocytes

Objective: Lower pregnancy rates of in vitro matured oocytes compared to those of invivo stimulated cycles indicate that optimization of in vitro maturation (IVM) remains achallenge. Reduced developmental competence of in vitro matured oocytes shows thatcurrent culture systems for oocyte maturation do not adequately support nuclear and/orcytoplasmic maturation. Therefore this study evaluates the effects of different concentrationsof saffron (Crocus sativus L.) aqueous extract (SAE), as an antioxidant agent on IVMof immature mouse oocytes.Materials and Methods: In this experimental study ,cumulus-oocyte complexes (COCs)were collected from 6-8 weeks old novel medical research institute (NMRI) female miceovaries. COCs were cultured in IVM medium supplemented with 0 (control), 5, 10, 20 and40 μg/ml of SAE in 5% CO2 at 37°C. The rates of maturation, fertilization and developmentwere recorded. ANOVA and Duncan’s protected least significant test, using the SASprogram was applied for all statistical analysis.Results: The maturation rate was significantly higher in all groups treated with differentconcentrations of SAE compared with the control group (p<0.05). However, the lowerconcentrations of SAE (10 and 5 μg/ml) in maturation medium respectively increased thefertilization rate of oocytes and in vitro developmental competence when compared withthe control group (p<0.05).Conclusion: The results of this study indicate that lower concentrations of SAE are moreappropriate to be added to maturation medium when compared with other experimental andcontrol groups. Generally, we conclude that addition of appropriate amounts of natural extractssuch as SAE to maturation medium improves oocyte maturation and embryo development