The impact of follicle-flushing during oocyte collection on embryo development of in-vitro fertilization.

BACKGROUND: To evaluate the impact of follicle-flushing during oocyte collection on embryo development potential retrospectively. METHODS: A total of 1714 cases, including 133 who experienced retrieval difficulty (repeated follicle-flushing) on the day of oocyte retrieval (difficulty group) and the control 1581 cases (control group), were assessed in this retrospective study. The number of oocytes recovered, two pro-nuclei fertilization (2PN-fertilization), day 3 good-quality embryo and day 5/6 blastocyst utilization rates were compared between the difficulty group and control group correspondingly. Embryo implantation, clinical pregnancy and neonatal outcomes were further analyzed between the two groups in the fresh day- 3 embryo transfer cycles. RESULTS: The number of oocytes recovered in the difficulty group (9.08 ± 4.65) were significantly reduced compared with the control group (12.13 ± 5.27),P < 0.001; The 2PN-fertilization, day 3 good-quality embryo and blastocyst utilization rates were significantly lower in the difficulty group compared with controls (71.7% vs. 75.7%; 52.7% vs. 56.5%; 31.9% vs. 37.0%, all P < 0.05). Embryo implantation in the difficulty group was 53.2%, which was lower than the control value of 58.7%, although not reaching statistical significance. The rate of fresh embryo transfer cycles in the difficulty group was lower than normal ones (51.88% vs. 61.99%, P = 0.026). The pregnancy and live birth rates were similar between the two groups. But the rate of spontaneous miscarriages of the difficulty group was higher than the control group, although not reaching statistical significance. The neonatal outcomes had no statistical difference between the two groups. CONCLUSIONS: Oocyte retrieval difficulty, which include repeated flushing and the corresponded extending time required for oocyte recovery, significantly reduced day 3 good-quality embryo and blastocyst utilization rates of these patients. But the live birth rate had no difference between the difficulty group and the normal ones.

[Repeated superovulation induction after failure in previous IVF-ET cycles with the ultra-long protocol: Analysis of outcomes of clinical pregnancy].

OBJECTIVE: To analyze the clinical outcomes of repeated superovulation induction in patients with adenomyosis or moderate to severe pelvic endometriosis after failure in previous IVF-ET cycles with the ultra-long protocol. METHODS: We retrospectively analyzed the clinical data about 37 patients with adenomyosis or moderate to severe pelvic endometriosis in our center from 2009 to 2013, who underwent repeated IVF-ET after failure in the previous cycles with the ultra-long protocol, namely by injection of 2－6 ampoules of 3.75 mg gonadotropin-releasing hormone agonist (GnRH-a). All the patients met the following requirements: hCG-negative at 14 days after transfer, within 3－7 days after menstruation, and properly down-regulated serum follicle stimulating hormone (FSH) (<10 mIU/ml), luteinizing hormone (LH) (<10 mIU/ml), estradiol (E2) (<30 pg/ml), follicle diameter (<10 mm) and endometrial thickness, and received GnRH (Gonal-F, Serono) for ovulation induction. We compared the clinical and laboratory data and pregnancy outcomes between the first and repeated cycles before and after ovulation induction. RESULTS: The repeated cycles, as compared with previous ones, showed significant increases in the antral follicle count (AFC) on the first day of stimulation (7.55 ± 1.86 vs 6.45 ± 2.5, P<0.05), number of follicles =≥14 mm in diameter on the hCG trigger day (7.81 ± 3.6 vs 5.56 ± 3.68, P<0.05), level of E2 (ï¼»2 362.15 ± 1 210.49ï¼½ vs ï¼»1 749.22 ± 1 139.44ï¼½ pg/ml, P<0.05), and numbers of oocytes retrieved (7.51 ± 3.23 vs 4.78 ± 3.41, P<0.05) and embryos transferred (2.00 ± 0.33 vs 1.50 ± 0.67, P<0.05), exhibited a remarkably reduction in the dose of GnRH (ï¼»1 791.65 ± 1 889.41ï¼½ vs ï¼»3 439.56 ± 1 836.53ï¼½ IU, P<0.05), and achieved a clinical pregnancy rate of 62.16%. CONCLUSIONS: With proper reduction of the FSH, LH and E2 levels and follicle diameter, repeated superovulation induction for IVF-ET can improve the ovarian response and pregnancy outcomes of the patients with adenomyosis or moderate to severe pelvic endometriosis after failure in the previous IVF-ET cycles with the ultra-long protocol.

FSH modulates the expression of inhibin-alpha and the secretion of inhibins via orphan nuclear receptor NUR77 in ovarian granulosa cells.

It has been previously reported that follicle-stimulating hormone (FSH) regulates the expression of inhibin-alpha in human granulosa cells, but the precise molecular pathway remains unknown. In the present study, we investigated the role of the orphan nuclear receptor, NUR77, in both the transcriptional regulation of the inhibin α-subunit gene and the secretion of inhibins. Our results showed that in a human granulosa cell tumor-derived cell line (KGN) and in human granulosa-lutein cells (hGL), FSH induced the expression of NUR77 and inhibin-alpha, although inhibin-alpha expression did not increased following FSH treatment if NUR77 was knocked down. Furthermore, simply overexpressing or reducing NUR77 levels affected inhibin-alpha expression, while NUR77 overexpression improved the secretion of inhibin A and B from human granulosa cells. In addition, chromatin immunoprecipitation-PCR, avidin-biotin-conjugated DNA precipitation, and luciferase reporter assays confirmed that NUR77 directly regulated the transcription of the inhibin-alpha gene through the specific NGFI-B response element located within its promoter. In the ovarian granulosa cells of the Nur77 knockout mice, the mRNA levels of inhibin-alpha were decreased relative to wild-type mice. These data indicate a role of NUR77 in the regulation of inhibin-alpha in ovarian granulosa cells.

Withdrawal of GnRH agonist decreases oestradiol and VEGF concentrations in high responders.

This study evaluated whether the withdrawal of a gonadotrophin-releasing hormone (GnRH) agonist before triggering ovulation reduces the incidence of ovarian hyperstimulation syndrome (OHSS) in high-risk infertility patients who were treated with gonadotrophins. GnRH agonist was withdrawn for 2 or 3 days when dominant follicles were ≥14 mm in diameter, according to the GnRH agonist long protocol. Non-withdrawal of GnRH agonist was used as control. The serum concentration of oestradiol on the ovulation trigger day was significantly decreased in the GnRH agonist withdrawal group compared with the control group (5750.78 ± 2344.77 pg/ml versus 8076.43 ± 1981.67 pg/ml); however, the number of retrieved oocytes and the fertilization rate were similar between the groups. In addition, the concentrations of vascular endothelial growth factor in plasma on day of human chorionic gonadotrophin administration and follicular fluid on the oocyte retrieval day were decreased following GnRH agonist withdrawal. In fresh embryo transfer cycles, rates of clinical pregnancy, implantation and OHSS were not different between the groups. When GnRH agonist withdrawal was followed by total embryos cryopreserved, the rate of OHSS was decreased compared with the control group (0% versus 8.70%). Clinical pregnancy rates in cryopreserved embryo transfer cycles were comparable between the two groups.

A case report of Trousseau syndrome.

RATIONALE: In 1865, Trousseau first discovered pulmonary embolism caused by multiple venous thrombosis in patients with gastric cancer, and later all clinical manifestations of malignant patients during pathogenesis due to abnormal coagulation and fibrinolysis were referred to collectively as Trousseau syndrome. Trousseau syndrome is not a benign thrombophlebitis, and when diagnosed it requires immediate treatment. The survival rate over 1 year is only 12%. Stroke in cancer patients has distinct characteristics different from conventional stroke and has higher mortality. PATIENT CONCERNS: A 54-year-old female presented to the Department of Otolaryngology with recurrent right nasal bleeding for 4 days. After surgery, the patient experienced 7 different cerebral infarction courses. Finally died of brain herniation. DIAGNOSIS: The specific abnormal laboratory index is the increase of D-dimer, suggesting the hypercoagulation state. The patient developed multiple cerebral infarction, myocardial injury, renal infarction, splenic infarction, and lower extremity arterial thrombosis, and finally was diagnosed Trousseau syndrome. INTERVENTIONS: In the treatment, aspirin and atorvastatin were selected, but it did not work very well. D-dimer were high, we used low molecular weight heparin, and D-dimer decreased significantly. OUTCOMES: Finally the patient died of brain herniation. CONCLUSION: The raise of D-dimer and typical magnetic resonance imaging manifestation which provides a greater basis for diagnosis. The specific abnormal laboratory index is the increase of D-dimer, which provides direction for treatment and helps to evaluate treatment effect.

[Incidence of polynuclear zygotes and pregnancy rate after short coincubation of gametes in in vitro fertilization].

OBJECTIVE: To explore the relationship of the incidence of polynuclear zygotes with clinical pregnancy after short coincubation of gametes in in vitro fertilization (IVF). METHODS: We retrospectively analyzed 3 862 cases of short gamete coincubation IVF, which were divided into six groups according to the percentage of polynuclear zygotes: 0,1-10%, 11-20%, 21-30%, 31-50%, 41-50%, and > or = 51%. We compared the rates of clinical pregnancy, implantation and abortion among the six groups. RESULTS: No statistically significant differences were found in the patients'age, dose of gonadotropin, peak E2, number of follicles at the hCG trigger, and number of oocytes retrieved among the six groups. The 1-10% group showed higher rates of pregnancy and implantation, while the > or = 51% group exhibited lower rates of pregnancy and implantation but a higher rate of abortion than the other groups, none with significant differences (P > 0.05). CONCLUSION: The incidence of polynuclear zygotes after short coincubation of gametes in IVF cannot serve as a prognostic indicator of the outcome of clinical pregnancy.

[Effectiveness of ICSI in patients with a high incidence of triploidy in a previous IVF cycle].

OBJECTIVE: To explore the effectiveness of ICSI in overcoming the high incidence of tripronucleates zygotes resulting from insemination in a previous IVF cycle. METHODS: We retrospectively analyzed the matched-pair cycles in 37 patients with a > 35 % incidence of tripronucleate zygotes in an IVF cycle, with ICSI used in the subsequent cycle, evaluated the incidences of diploid (2PN) and triploid (3PN) zygotesand the number of normal embryos obtained, and compared the rates of clinical pregnancy and embryo implantation between the IVF and ICSI groups. RESULTS: The mean age of the ICSI group was significantly older than that of the IVF group, while the ampules of gonadotropin and peak E2 showed no remarkable difference between the two. The numbers of follicles at hCG trigger, retrieved oocytes and mature oocytes were markedly lower in the former than in the latter. The percentage of 2PN was significantly higher while that of 3PN significantly lower after ICSI than after IVF (74.24% vs 34.42%; 11.57% vs 51.04%, P < 0.01), and more normal diploid embryos were obtained with ICSI (3.83 +/- 2.08 vs 2.52 +/- 1.71, P < 0.01). Four singletons were achieved in 31 IVF embryo transfer cycles, in comparison with 11 singletons and 3 twins in 36 ICSI embryo transfer cycles. The ICSI group showed significantly higher rates of clinical pregnancy and embryo implantation than the IVF group (38.89% vs 12.90%; 28.33% vs 7.41%, P<0.01). CONCLUSION: For women with a high incidence o triploidy in a previous IVF cycle, ICSI can effectively increase the number of normal diploid zygotes.

[Pregnancy outcomes of repeated cycles of in vitro fertilization and embryo transfer].

OBJECTIVE: To analyze the pregnancy outcomes of repeated IVF-ET cycles. METHODS: We retrospectively analyzed 702 repeated IVF-ET cycles (503 cases) performed in our center from 2006 to 2008, among which 191 cycles (Group A) had failed previously in other hospitals and 511 (Group B) in ours, focusing on the relationship of pregnancy outcomes with the number of repeated IVF-ET cycles and the age of the patients. RESULTS: In Group A, there were no significant differences in pregnancy rates among the patients with 1, 2 or > 2 previously failed cycles (56.56% vs 66.67% vs 61.54%), while the numbers of oocytes obtained were significantly decreased (8.51 +/- 4.60 vs 8.48 +/- 3.32 vs 4.86 +/- 2.96) and the serum levels of follicle stimulating hormone (FSH) remarkably increased ([7.31 +/- 3.66] mIU/mL vs [6.83 +/- 2.35] mIU/mL vs [11.58 +/- 11.40] mIU/mL) in those with > 2 previous failures. In Group B, the results of the first IVF-ET cycle in our center showed that the number of oocytes obtained and the E2 level on the day of hCG injection were markedly decreased in the patients with 2 previously failed cycles (6.66 +/- 4.58 vs 9.59 +/- 4.30 and 2 396.87 +/- 1 602.02 vs 4 061.17 +/- 2 255.63), and so were the pregnancy rate, oocyte number, intimal thickness and essential FSH level in those with > 2 previous failures, but no significant differences were found in the rate of pregnancy, number of oocytes obtained, number of embryos transplanted and rate of abortions in those with 1 previous failure. The pregnancy and implantation rates of the repeated IVF-ET cycles were significantly reduced in the female patients aged < 38 years and with > 3 previously failed cycles, as well as in those aged > 38 years and with 1 - 4 previous failures, but not in those aged < 38 years and with < 3 previous failures in Group B. CONCLUSION: The pregnancy outcome of repeated IVF-ET cycles was not correlated with the number of the cycles, but maybe directly with different protocols in different reproductive centers. The rate of pregnancy was obviously decreased in patients that underwent over 4 repeated IVF-ET cycles, but had no obvious correlation with the number of cycles in those that received 1 - 3 cycles in the same reproductive center. The age of the patient influences the results of repeated IVF-ET cycles, and both pregnancy and implantation rates may decrease in those aged > 38 years.

[Early rescue intracytoplasmic sperm injection: safe for complete IVF failure].

OBJECTIVE: To evaluate the safety of early rescue intracytoplasmic sperm injection (ICSI) for complete failure of in vitro fertilization (IVF). METHODS: We conducted early rescue ICSI for 105 conventional IVF cycles and follow-up evaluation of the clinical outcomes. RESULTS: A total of 64 neonates were born, and no significant differences were found in the sex ratio, birth weight and birth defects. CONCLUSION: Early rescue ICSI can significantly improve the clinical outcome in patients with complete IVF failure, but does not increase the clinical risk.

[Effects of fertilization methods and sperm sources on the developmental capacity of surplus embryos].

OBJECTIVE: To explore the effects of fertilization methods and sperm sources in intracytoplasmic sperm injection (ICSI) on the developmental capacity of surplus embryos. METHODS: We analyzed the blastocyst formation of the surplus embryos from 2 135 patients, who were divided according to fertilization methods into an IVF (n=1803) and an ICSI group (n=332), the former again allocated to a normal fertilization (n=1642) and a rescue fertilization group (n=161), and the latter, according to sperm sources, to an ejaculated (n=248), an epididymal (n=70) and a testicular sperm group (n=14). The rates of blastocyst formation and good-quality blastocysts were compared between different fertilization methods and sperm sources. RESULTS: A total of 1884 blastocysts (28.87%) formed from 6525 surplus embryos of the patients after sequential culture, of which 974 (51.70%) were good-quality ones. The blastocyst formation rate of surplus embryos was significantly higher in the IVF (30.14%) than in the ICSI group (21.40%, P < 0.05), the rate of good-quality blastocysts was also higher in the former (52.44%) than in the latter (45.54%), but with no significant difference (P > 0.05). The rates of blastocyst formation and good-quality blastocysts were significantly higher in the normal (31.04% and 53.28%) than in the rescue fertilization IVF group (20.38% and 38.54%, P < 0.05), and in the testicular sperm ICSI group (30.23% and 53.85%) than in either the epididymal (18.36% and 42.11%) or the ejaculated sperm ICSI group (21.76% and 45.70%) (P < 0.05). CONCLUSION: The development potential of surplus embryos was higher in IVF than in ICSI, in the normal than in the rescue fertilization IVF group, and in the testicular than in the epididymal and ejaculated sperm ICSI groups.

[Establishment of a method for quantifying tuberous sclerosis complex 2 mRNA in mouse single oocyte and preimplantation embryo].

OBJECTIVE: To develop a method for quantifying gene expressions in the mouse single oocyte and preimplantation embryo. METHODS: We quantified the message RNA (mRNA) expression of the TSC2 gene in the single oocyte and preimplantation embryo by capillary electrophoresis using the exogenic mutation TSC2 gene as the reference and amplification by competition polymerase chain reaction (PCR). RESULTS: We successfully established the method for quantifying the mRNA expression of the TSC2 gene, with good linear relations between the mRNA level of the TSC2 gene and the dilution degree of the reference gene (r = -0.987). CONCLUSION: The level of the mouse TSC2 gene expression can be effectively quantified by competition PCR and capillary electrophoresis, which has provided a molecular base for evaluating the quality of human oocytes and preimplantation embryos.

[Efficient treatment of severe teratozoospermia by ICSI with testicular spermatozoa: a report of 5 cases].

OBJECTIVE: To explore the feasibility of treating severe teratozoospermia (the abnormity rate of ejaculated or epididymal sperm > or = 99%) by intracytoplasmic sperm injection (ICSI) with testicular sperm so as to improve the outcome of assisted reproductive technology. METHODS: We retrospectively analyzed the clinical data of 5 patients with severe teratozoospermia (epididymal: n=4; ejaculated: n=1) treated by ICSI with sperm from different sources, and compared the rates of fertilization, cleavage, quality embryos, pregnancy and implantation between the testicular and non-testicular sperm groups. RESULTS: Four ongoing clinical pregnancies were achieved by ICSI with testicular sperm, but none with ejaculated (or epididymal) sperm. The rates of pregnancy and implantation were significantly higher in the testicular than in the non-testicular sperm group (P < 0.01), and there were no significant differences in the rates of fertilization, cleavage and quality embryos between the two groups (P > 0.05). CONCLUSION: ICSI with testicular sperm could efficiently improve the therapeutic outcome for men with severe teratozoospermia.

[Combination of short-period sperm-oocyte coincubation and early rescue intracytoplasmic sperm injection after total failure of in vitro fertilization].

OBJECTIVE: To evaluate the outcome of the combination of short-period sperm-oocyte coincubation with early rescue intracytoplasmic sperm injection (ICSI) after complete failure of in vitro fertilization (IVF). METHODS: We used short-period sperm-oocyte coincubation and overnight fertilization for the oocytes obtained from 10 IVF-ET cycles, and compared the fertilization results and embryo quality. We conducted polar body observation following short-period sperm-oocyte coincubation for 105 conventional IVF cycles, and evaluated the clinical outcome of early rescue ICSI for total fertilization failure. RESULTS: No significant differences were noted in fertilization outcome and embryo quality between the 3 h short-period and overnight fertilizations (P > 0.05). The combined strategy achieved a clinical pregnancy rate of 53.3%, an implantation rate of 38.0%, with 64 babies born, of whom 44.8% were taken home. CONCLUSION: The combination of short-period sperm-oocyte coincubation with early rescue ICSI can significantly improve the clinical outcome in patients who have experienced complete failure of in vitro fertilization.

[Comparison of 3 techniques for artificial shrinkage of blastocoelic cavity before vitrification of expanded mouse blastocysts].

OBJECTIVE: To establish an effective pretreatment method for the vitrification of expanded mouse blastocysts by comparing 3 techniques for the artificial shrinkage of the blastocoelic cavity. METHODS: The blastocoelic cavity was artificially shrunk by micro-needle aspiration, pipetting or laser drilling prior to the vitrification of the expanded blastocysts. The rates of survival and hatching achieved with the three techniques were compared with those of the non-shrinkage group. RESULTS: The rates of survival were 72.9, 72.0 and 94.0%, and those of hatching were 64.6, 32.0 and 62.0% in the three shrinkage groups, obviously higher than in the nonshrinkage group (40.0 and 16.0%). CONCLUSION: Artificial shrinkage of the blastocoelic cavity was an effective pretreatment technique for the vitrification of expanded mouse blastocysts, especially by micro-needle aspiration and laser drilling.

[Sperm DNA damage and assisted reproductive technology].

With the introduction of assisted reproductive technology (ART), sperm assessment has developed progressively, from conventional semen routine tests to novel cellular and molecular measures. Sperm DNA damage is a new marker of male fertility, whose genetic mechanism involves abnormal package and segregation of chromatin, oxidative stress, abnormal cell apoptosis, etc. Sperm chromatin structure assay (SCSA) is one of the common techniques to measure sperm DNA damage. Sperm DNA damage might be associated with the pregnancy outcome of ART, recurrent spontaneous abortion and potential genetic risk of ICSI offspring. Some treatment strategies might reduce the percentage of sperm DNA damage and increase the success rate of ART, including oral administration of antioxygen drugs, ICSI with testis sperm, sperm freezing and preservation, removing of etiological factors, traditional Chinese medicine, and so on. This review focuses on the mechanism and detection of sperm DNA damage, its association with reproductive outcomes, and relevant treatment strategies in assisted reproductive technology.

[Application of GnRH-antagonist to IVF-ET for patients with poor ovarian response].

OBJECTIVE: To compare the outcomes achieved by GnRH-antagonist and GnRH-agonist in IVF-ET for patients with poor ovarian responses, and to find out a better protocol for ovulation stimulation. METHODS: (1) Patients with poor ovarian responses were assigned to an experimental (n = 63) and a control group (n = 58), treated respectively with GnRH-Ant and oral contraceptive plus micro-dose GnRH-a (OC + GnRH-a), and comparisons were made of the medication doses, laboratory results and pregnancy outcomes between the two groups. (2) Twenty of the patients were treated first with GnRH-Ant and then with OC + GnRH-a, and the same comparisons were made between the two protocols. RESULTS: Between the experimental and the control groups, there were no significant differences in the dose of Gn and number of retrieved oocytes and transplanted embryos (P > 0.05), nor in the pregnancy rate of transplantation cycles (37.29% vs 35.29%). The cycle cancellation rate was lower in the experimental than in the control group (6.35% vs 12.07%), with no statistical difference (P > 0.05). The cycle duration was significantly different between the two groups ([9.65 +/- 1.60] d vs [19.05 +/- 3.94] d) (P < 0.05). As for the comparison of GnRH-Ant with GnRH-a, no significant differences were observed in the dose of Gn and the numbers of retrieved oocytes and transplanted embryos (P > 0.05). GnRH-Ant achieved a higher pregnancy rate of transplantation cycles but a significantly lower cancellation rate than GnRH-a (38.09% vs 17.64% and 0% vs 15%) (P > 0.05 and P < 0.01), respectively. The cycle duration of the former was statistically shorter than that of the latter ([9.91 +/- 2.49] d vs [27.74 +/- 25.39] d) (P < 0.05). CONCLUSION: Compared with micro-dose GnRH-a, GnRH-Ant can shorten the cycle duration and reduce the cancellation rate in IVF-ET for patients with poor response. And for those who have failed to respond to GnRH-a, GnRH-Ant may be tried in another attempt at IVF-ET.

[Establishment of human embryonic stem cell line NJGLLhES1].

OBJECTIVE: To report the derivation and characterization of a new human embryonic stem cell (hESC) line NJGLLhES1. METHODS: From the inner cell mass of frozen-thawed human embryos and with ICR mouse embryonic fibroblasts as the feeder layer, we established a new human embryonic stem cell line, which was named NJGLLhES1. We detected the karyotype of the cell line, determined the expressions of alkaline phosphatase, the specific cell surface antigens SSEA-3, SSEA-4, TRA-1-60, TRA-1-81 and the marker gene Oct-4, and examined the formation of embryoids and teratomas. RESULTS: NJGLLhES1 was maintained for over 1 year in vitro, with the morphological characteristics of hESC, a normal karyotype, positive expressions of alkaline phosphatase and specific cell marker genes, and the potential of forming embryoids and teratomas. CONCLUSION: A new human embryonic stem cell line NJGLLhES1 was successfully established, which remains karyotypically and phenotypically stable, undifferentiated and capable of self-renewal and pluripotential differentiation.

[Laser-assisted immobilization causes no direct damage to sperm DNA].

OBJECTIVE: To determine whether laser-assisted immobilization of sperm damages sperm DNA. METHODS: Twenty-three semen samples were selected from an IVF program. Then normal spermatozoa were obtained by swimming-up method and immobilized with the tail by 0.45 ms pulse laser. Terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) and single cell gel electrophoresis (SCGE) were used to detect sperm DNA damage. RESULTS: There was no significant difference either before and after laser treatment in the percentage of TUNEL-positive spermatozoa ([1.32 +/- 0.61]% vs [1.41 +/- 0.51]%, P > 0.05) or in SCGE ([1.59 +/- 0.70]% vs [1.83 +/- 0.68]%, P > 0.05). CONCLUSION: Laser-assisted sperm immobilization may cause no direct damage to the sperm DNA.

[The application of quality control system in reproductive medicine center].

Uniform standards are required for evaluating the quality of reproductive medicine center engaged in vitro-fertilization and embryo transfer. Clinic pregnancy rates in common use are not in evaluation focus. More internationalized standard methods should be used to assess the quality of IVF center.

[Establishment of autologous endometrial coculture and sequential system for human early embryo culture].

OBJECTIVE: To establish a coculture and a sequential system for human early embryo culture. METHODS: The endometrial tissue was digested enzymatically and cultured to achieve generated and cryo-thawed endometrial monolayer cells. The generated and cryo-thawed monolayer cells were cocultured with human 2PN embryos and transferred to sequential medium every 48 hours. RESULTS: Human endometrial cells had viability in vitro culture. The autologously generated and cryo-thawed monolayer cells were successfully obtained, and 74.04% of the cryo-thawed cells were successfully used in coculturing human early embryos. The embryos developed well, with the clinical pregnancy rate of 68.83% and the implantation rate of 44.23%. CONCLUSION: The autologous endometrial cell coculture and sequential culture system for human early embryo development provides a feasible method for studying human embryo development and implantation so as to improve embryo quality.