Cumulus Cells Are Potential Candidates for Cell Therapy.

BACKGROUND/AIM: Cumulus cells (CCs) originate from the membrane granulosa cells and surround oocytes during follicle maturation. CCs produce high levels of hyaluronan that targets CD44, which is a major tumorigenic marker. This study aimed to investigate whether CCs have a role in cell therapy by targeting CD44 in pancreatic cancer cells. MATERIALS AND METHODS: CCs were isolated from the oocytes and incubated in a hypoxic environment. BxPC-3 pancreatic cancer cells were treated with CC conditioned media for three days. RESULTS: Conditioned media of CC cells incubated in hypoxic conditions caused a 25% reduction in the viability of BxPC-3 cells. Expression of anti-apoptotic genes was down-regulated, while that of pro-apoptotic genes was upregulated. An increased number of BxPC-3 cells exhibited increased levels of reactive oxygen species and arrested in the synthesis (S) phase of the cell cycle. CONCLUSION: CCs conditioned medium induced apoptosis of pancreatic cancer cells.

The cutoff values of serum AMH levels and starting recFSH doses for the individualization of IVF treatment strategies.

OBJECTIVE: The main purpose of our study is to categorize starting doses of recombinant follicle-stimulating hormone (recFSH) based on various cutoff values of anti-Mullerian hormone (AMH) and to determine the effectiveness of serum AMH levels in the prediction of poor ovarian response. MATERIAL AND METHODS: Prospective data analysis was conducted at IVF center. A total of 323 patients were included. All patients were divided into four groups according to the patients' serum AMH concentrations: Group 1 (AMH < 1 ng/ml; 450 IU/day n = 157); Group 2 (AMH 1-2 ng/ml; 375 IU/day, n = 55); Group 3 (AMH 2-3 ng/ml; 225 IU/day, n = 48); and Group 4 (AMH > 3 ng/ml; 150 IU/day, n = 63). Collected data included age, total gonadotropin dosage, duration of stimulations, the total number of oocytes retrieved, ovarian response, cancelation rate, and cPRs. RESULTS: As serum AMH levels increased, there were significant decreases in the starting recFSH dose and total gonadotropin dosage, and a significant increase in the total number of oocytes retrieved. There was a significant trend toward increasing cycle cancelation rates and decreasing cPRs with decreasing serum AMH levels. Although there were no significant differences with regard to the proportion of cycles with hypo-response between all groups. A result of ≤0.83 was considered the cutoff value of AMH to predict a hypo-response to ovarian stimulation. CONCLUSIONS: AMH is a useful marker in selecting the starting dose of recFSH and prediction of poor ovarian response. Our protocol may allow clinicians to modulate the starting dose of recFSH according to these cutoff values for serum AMH levels.

The importance of the length of uterine cavity, the position of the tip of the inner catheter and the distance between the fundal endometrial surface and the air bubbles as determinants of the pregnancy rate in IVF cycles.

OBJECTIVE: To evaluate the relationship between the pregnancy rate (PR) and the positioning of the intrauterine catheter at embryo transfer (ET) under transabdominal ultrasound (US) guidance in in vitro fertilization (IVF) cycles. STUDY DESIGN: Prospective data analysis of 281 consecutive US-guided fresh ETs performed by a single physician at Yeditepe University Hospital IVF Center, Istanbul, Turkey, after controlled ovarian hyperstimulation between April 2012 and March 2013. The length of the uterine cavity (A), the distance between the fundal endometrial surface and the tip of inner catheter (B), the distance between the fundal endometrial surface and the air bubbles (C), and the pregnancy rates (PRs) were recorded. RESULTS: The mean age of the patients was 33.25±5.5 years. Of all transfers, 115 (40.9%) resulted in a clinical pregnancy. With regard to distance (C), the clinical intrauterine pregnancy rates were 65.2%, 32.2% and 2.6% in the <10mm, 10-20mm, and 20mm distance groups, respectively. The PR was dramatically reduced in cases with >10mm between the fundal endometrial surface and the air bubbles, although this did not reach statistical significance. Between those patients who conceived and those who did not, there was no significant difference in terms of the distance between the fundal endometrial surface and the tip of inner catheter, the ratio of A/B or the ratio of B/C. CONCLUSIONS: The final position of the air bubble used as an identifier of the position of the embryo at ET can be determinative for PR, although it cannot be predicted. Clinical pregnancy rates appeared higher in cases with air bubbles closer to the fundus and the optimal position of the air bubble seems to be a distance of <10mm from the fundal endometrial surface. It could be advisable to monitor the final position of air bubble at ET for identifying PR. In addition, the depth of uterine cavity may be considered to indirectly be important factor as it affects ET depth. The optimal distance between the fundal endometrial surface and the tip of inner catheter is 1.5-2cm. Further well-designed randomized controlled trials are required to optimize ET technique in the future.

Comparison of follicular fluid and serum cytokine concentrations in women undergoing assisted reproductive treatment with GnRH agonist long and antagonist protocols.

BACKGROUND: This study investigated whether follicular fluid (FF) and serum (S) concentrations of cytokines in women undergoing assisted reproductive treatment (ART) were different in GnRH antagonist cycles compared to agonist long ones. METHODS: A retrospective clinical study was performed at a University ART center. A total of 85 women who underwent ART either with agonist long (n = 34) or antagonist protocol (n = 51) were analyzed. FF and serum samples were collected at the time of oocyte retrieval and measured for interleukin (IL)- 1beta, IL-6, IL-8, IL-12, tumor necrosis factor (TNF)-alpha by the enzyme-linked immunosorbent assay (ELISA) technique, using commercially available kits and nitric oxide (NO) by the nitrate/nitrite colorimetric assay. The results were compared between GnRH antagonist and agonist cycles. RESULTS: No significant difference was found in the FF concentrations of those cytokines between the two protocols. The serum values were also similar in the two groups except IL-6 (14.3 +/- 4.8 vs. 20.5 +/- 12.2 pg/ml; p = 0.008) and NO (1.4 +/- 1.1 vs. 2.2 +/- 1.9 microm; p = 0.038) levels which were found to be significantly lower in antagonist cycles. CONCLUSIONS: There is no significant difference in follicular microenvironment in terms of IL-1beta, IL-6, IL-8, IL-12, TNF-alpha, and NO levels between agonist long and antagonist cycles. However, serum IL-6 and NO levels were lower in women given antagonists.

Can pentoxifylline improve the sperm motion and ICSI success in the primary ciliary dyskinesia?

Primary ciliary dyskinesia (PCD), previously known as immotile cilia syndrome, can cause respiratory and reproductive problems. Because of the impaired motion of microtubules the patients suffer upper respiratory tract problems. Infertility is an other issue of these patients. Several attempt have been proposed to bring back sperm motion and to improve intracytoplasmic sperm injection (ICSI) results. With our case we have described the management of a male infertility that causes from immotile cilia. The role of ICSI with incubated and activated ejaculatory sperm by pentoxifylline in the patient of PCD or as commonly named immotile cilia syndrome.

Cytokine and nitric oxide concentrations in follicular fluid and blood serum of patients undergoing assisted reproductive treatment: relationship to outcome.

OBJECTIVE: The role of cytokines and nitric oxide (NO) in ovarian folliculogenesis and the development of mature and fertilizable oocytes is controversial. The aim of this study is to determine the concentrations of interleukin (IL)-1ß, IL-6, IL-8, IL-12, tumor necrosis factor (TNF)-α and NO in the follicular fluid (FF) and blood serum (S) of patients undergoing assisted reproductive treatment (ART) and to investigate whether these cytokines could be used as a predictive parameter for ART outcome. MATERIAL AND METHODS: A retrospective clinical study was performed at a university hospital including a total of 85 women who underwent ART. FF and serum samples were collected at the time of oocyte retrieval and measured for interleukin (IL)-1ß, IL-6, IL-8, IL-12, tumor necrosis factor (TNF)-α by the enzyme-linked immunosorbant assay (ELISA) technique, using commercially available kits and NO by the nitrate/nitrite colorimetric assay. The results were compared between the women who became pregnant and those who did not following ART. RESULTS: No significant difference was found in the FF and blood serum concentrations of the cytokines and NO between pregnant and non-pregnant women. CONCLUSION: Follicular fluid and blood serum concentrations of IL-1ß, IL-6, IL-8, IL-12, TNF-α and NO do not predict pregnancy achievement following ART.

Establishment and characterization of new human embryonic stem cell lines.

Human embryonic stem cells (hESC), with their ability to differentiate into all cell types in the human body, are likely to play a very important therapeutic role in a variety of neurodegenerative and life-threatening disorders in the near future. Although more than 120 different human embryonic stem cell lines have been reported worldwide, only a handful are currently available for researchers, which limits the number of studies that can be performed. This study reports the isolation, establishment and characterization of new human embryonic stem cell lines, as well as their differentiation potential into variety of somatic cell types. Blastocyst-stage embryos donated for research after assisted reproductive techniques were used for embryonic stem cell isolation. A total of 31 blastocysts were processed either for immunosurgery or direct culture methods for inner cell mass isolation. A total of nine primary stem cell colonies were isolated and of these, seven cell lines were further expanded and passaged. Established lines were characterized by their cellular and colony morphology, karyotypes and immunocytochemical properties. They were also successfully cryopreserved/thawed and showed similar growth and cellular properties upon thawing. When induced to differentiate in vitro, these cells formed a variety of somatic cell lineages including cells of endoderm, ectoderm and mesoderm origin. There is now an exponentially growing interest in stem cell biology as well as its therapeutic applications for life-threatening human diseases. However, limited availability of stem cell lines as well as financial or ethical limitations restrict the number of research projects. The establishment of new hESC lines may create additional potential sources for further worldwide and nationwide research on stem cells.

The effects of a caries-removing gel (Carisolv) on FM3A cell-line in vitro.

OBJECTIVES: The aim of this study was to examine the cytotoxic effects of Carisolv on mouse mammary carcinoma cell line (FM3A) at different application times. METHODS: The FM3A cell line obtained from the European Collection of Animal Cell Cultures was used in the cell culture assays. Exponentially growing cells were seeded in 5x10(5)cells/well in 5 ml of RPMI 1640 medium supplemented with 10(%) fetal calf serum and antibiotics in each well of a six-well plate. Carisolv gel was applied onto the cell culture medium for 1, 5 and 20 min and incubated for 24 h at 37 degrees C in a humidified atmosphere of 5(%) carbon dioxide (CO(2)). After 24 h incubation, the cells were collected by trypsinisation and counted with a hemocytometer. The cytotoxicity of the Carisolv was determined by evaluation of cell growth and viability in comparison to untreated controls (cell growth=100%). For cell viability, the trypane blue exclusion assay was used. Dunnett's t-test was used for statistical analysis. RESULTS: Cell growth was significantly reduced after 20 min application of Carisolv in comparison to the control (p < 0.001) and 1 min treatment groups (p < 0.05) No significant differences were found in cell viability between the study groups. CONCLUSIONS: It can be concluded that prolonged application of Carisolv did not affect cell viability, but had a reducing effect on cell growth in the FM3A cell line.