Study question is there correlation between the mtDNA copy number in cumulus cells and anti-Mullerian hormone level,female age, oocyte quality, embryo morphology, ploidy and blastocyst implantation rate? Summary answer A positive correlation of mtDNA quantity in CCs with the AMH,female age was revealed.There was no correlation between mtDNA quantity and embryo morphology,ploidy,implantation rate What is known already Recent studies have suggested that age-related decreased competence of oocytes may be due to low quantity of mtDNA copy number. Moreover, quantification of mtDNA in CCs may serve as an predictor of blastocysts viability. The purpose of this study is to investigate relative levels of mtDNA in the OCCCs in association with female age, ovarian reserve, embryo morphology, ploidy and blastocyst implantation rate. Study design, size, duration Prospective clinical study performed on 470 CCs retrieved from 72 advanced reproductive age patients undergoing ART treatment with intracytoplasmic sperm injection (ICSI) and preimplantation genetic testing for aneuploidy (PGT-A). Out of the 130 obtained blastocysts 56 embryos were diagnosed as aneuploid, and 74 as euploid. Presently, 51 frozen euploid embryos were transferred (FET). All transferred euploid blastocysts (n = 51) divided into 2 groups: 1 group (n = 21) - implanted embryos, 2 group (n = 30) - non-implanted. Participants/materials, setting, methods Inclusion criteria: age 35-45years;BMI: 18 – 24,9kg/m2; FSH ≤15IU/ml; normal female/male karyotype; non-smokers. Exclusion criteria: genital endometriosis III-IV;severe extragenital pathology; polycystic ovary syndrome; chronic endometritis; > 96% of sperm with abnormal morphology (WHO criteria). MtDNA was assessed by using a quantitative real-time polymerase chain reaction technique. DNA from the trophectoderm samples were amplified and subjected to aneuploidy analysis using array comparative genomic hybridization. In statistic analysis was used Pearson’s correlation coefficients and Fisher’s exact test; p < 0.05 was considered significant. Main results and the role of chance The median age was 37.8 years old (range, 35–45), the mean level of anti-Mullerian hormone (AMH) was 2.66±1.09, and the mean body mass index was 22.3±1.5. A positive correlation of the relative level of mtDNA in the CCs with the patients' age (p = 0.008) and AMH levels (p = 0.003) was revealed. There was no statistically significant correlation between mtDNA copy number and embryo morphology on day 5 (p = 0.7). There was a tendency to increase mtDNA copy number in group 1 vs. group 2, 390 and 299, respectively (p > 0.05). In this study we didn’t find relationship between median mtDNA content of CCs and embryos ploidy (356 vs. 325, in euploid (n = 74) and aneuploidy (n = 56) blastocyst, respectively, p > 0.05). Limitations, reasons for caution Our study was carried out in a relatively small subset of participants and obtained embryos. Correspondingly, the results obtained cannot be readily extrapolated on other groups of patients and need to be confirmed in larger trials. Wider implications of the findings This study showed that mtDNA quantification in CCs isn’t a useful biomarker for prediction embryos implantation potential or ploidy. MtDNA content in CCs correlated only with female age and AMH level. Trial registration number *
Study question Does the use of granulocyte colony-stimulating factor (G-CSF) and platelet-rich (PRP) plasma improve endometrial thickness and pregnancy outcome in patients with thin endometrium and RIF? Summary answer The use of G-CSF and PRP improve the endometrial thickness and pregnancy outcome in patients with thin endometrium and RIF undergoing artificial FET cycles What is known already Thin endometrium is a negative prognostic factor for achieving pregnancy, which occurs in 2.4% of assisted reproductive technology cycles. According to previous studies use of G-CSF and PRP are perspective methods for these patients’ management. G-CSF is a polypeptide that belongs to the colony-stimulating factor glycoprotein group. PRP is derived from patient’s own blood and contains platelets’ growth factors that promote reparation in tissues. However, there is limited data about IVF treatment success after G-CSF or PRP perfusion in patients with thin endometrium and RIF, also there are no studies comparing these two protocols. Study design, size, duration This prospective observational study included 163 patients. All the patients underwent artificial frozen-thawed embryo transfer cycles. Group 1 (n = 43), the control group, did not get additional therapy. Group 2 (n = 46) received intrauterine G-CSF (300 mcg) perfusion on 5-6 and 8-9 days of cycle. Group 3 (n = 74) received intrauterine PRP (5-7 ml) perfusion on 8-9, 10-11 and 12-13 days of cycle. The study was carried out between December 2017 and December 2019. Participants/materials, setting, methods Inclusion criteria: age 20-42 years; BMI 18-30 kg/m2; endometrial thickness on embryo transfer (ET) day ≤ 7mm in previous cycles; previous non-effective ET cycles; good quality embryos (Gardner and Schoolcraft classification). Exclusion criteria: uterine fibroids ≥4 cm; deep endometriosis; Asherman syndrome; allergic reaction to G-CSF. PRP was prepared by the double centrifugation technique (Beckman, USA). From 400 ± 50 ml of whole blood was made 40 ± 5 ml PRP with 0,6-0,7 x 1011 platelets. Main results and the role of chance Endometrial thickness on ET day in Group 1 was 7,0±1,1 mm, in Group 2 – 7,9±1,8 mm, in Group 3 – 8,0±1,3 mm. The difference was statistically significant between Group 1 and 2 – p = 0,003 (MD-0,96; 95% CI: 0,33; 1.59), between Group 1 and 3 – p = <0,001 (MD-1,0; 95% CI: 0,53; 1,47). No statistically significant difference was between Group 2 and 3 – p = 0,790 (MD-0,004; 95% CI: -0,53; 0,61). FET cycle cancelled due to thin endometrium occurred in 26 (60,5%), 18 (39,1%) and 13 (17,6%) patients in Groups 1,2 and 3 respectively. Pregnancy rate was statistically significant different between Group 1 and 2 – p = 0,005 (5,9% (1/17) vs. 46,4% (13/28), OR-0,07; 95%CI: 0,01; 0,62), between Group 1 and 3 – p = 0,007 (5,9%(1/17) vs. 41,0%(25/61), OR-0,09; 95%CI: 0,01; 0,72). No statistically significant difference was between Group 2 and 3 in pregnancy rate – p = 0,630 (46,4% (13/28) vs. 41,0% (25/61)). There was statistically significant difference in livebirth rate between Group 1 and 2 – p = 0,003 (0 (0/17) vs. 39,3% (11/28), between Group 1 and 3 – p = 0,014 (0 (0/17) vs. 27,9% (17/61). No statistically significant difference was in livebirth rate between Group 2 and 3 – p = 0,282 (39,3% (11/28) vs. 27,9% (17/61)). Limitations, reasons for caution Our study was carried out in a relatively small subset of patients; also in our study we investigated only FET cycles. There should be larger trials in future, including fresh embryo transfer cycles. Wider implications of the findings This study opens new possibilities in management of patients with thin endometrium. Trial registration number N/A
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