CD317+ MSCs expanded with chemically defined media have enhanced immunological anti-inflammatory activities.

BACKGROUND: Although both preclinical and clinical studies have shown the great application potential of MSCs (mesenchymal stem/stromal cells) in treating many kinds of diseases, therapeutic inconsistency resulting from cell heterogeneity is the major stumbling block to their clinical applications. Cell population diversity and batch variation in the cell expansion medium are two major inducers of MSC heterogeneity. METHODS: Cell population diversity was investigated through single-cell RNA sequencing analysis of human MSCs derived from the umbilical cord and expanded with fully chemically defined medium in the current study. Then, the MSC subpopulation with enhanced anti-inflammatory effects was studied in vitro and in vivo. RESULTS: Our data showed that MSCs contain different populations with different functions, including subpopulations with enhanced functions of exosome secretion, extracellular matrix modification and responses to stimuli (regeneration and immune response). Among them, CD317+ MSCs have improved differentiation capabilities and enhanced immune suppression activities. Underlying mechanism studies showed that higher levels of TSG6 confer enhanced anti-inflammatory functions of CD317+ MSCs. CONCLUSIONS: Thus, CD317+ MSCs might be a promising candidate for treating immunological disorder-related diseases.

Hormone replacement therapy with GnRH agonist pretreatment improves pregnancy outcomes in patients with previous intrauterine adhesions.

PURPOSE: To explore the optimal cycle regimen for frozen-thawed embryo transfer (FET) in women with previous intrauterine adhesions (IUAs). METHODS: In this retrospective cohort study, a total of 1,002 FET cycles for patients with previous IUAs from January 2015 to December 2020 were included. Among them, 294 conventional hormone replacement therapy (HRT) cycles were matched with 155 HRT with gonadotropin releasing hormone agonist pretreatment (HRT+GnRH-a) cycles using propensity score matching. Multivariate logistic regression analysis was performed to further investigate the impact of cycle regimen on pregnancy outcomes. RESULTS: After propensity score matching, baseline characteristics were consistent between HRT and HRT+GnRH-a group. Logistic regression analysis revealed that there was a significant superiority of HRT+GnRH-a over the conventional HRT group regarding the incidences of live birth (aOR=1.966, 95%CI: 1.212-3.188, P=0.006) and ongoing pregnancy (aOR=1.710, 95%CI: 1.057-2.767, P=0.029). HRT+GnRHa also had a higher odd of clinical pregnancy (aOR=1.414, 95%CI: 0.903-2.216, P=0.130), and lower odd of early miscarriage (aOR=0.511, 95%CI: 0.219-1.195, P=0.121) compared to HRT, yet not reached statistical significance. CONCLUSION: HRT with GnRH-a pretreatment improves pregnancy outcomes in women with previous IUAs. GnRH-a may restore the endometrial receptivity in the FET cycles of IUAs. SYNOPSIS: HRT with GnRH-a pretreatment may promote pregnancy prognosis of FET in women with history of IUAs by restoring endometrial receptivity.

Endometriosis is associated with a lowered cumulative live birth rate: A retrospective matched cohort study including 3071 in vitro fertilization cycles.

Endometriosis, defined as the presence of endometrial tissues outside the uterus, affects approximately 10% of women of reproductive age. Assisted reproductive technology (ART) is considered the most effective technique to treat infertility in women, although the impact of endometriosis on ART outcomes remains ambiguous. In this study, 433 patients with endometriosis and 1299 infertile patients with tubal factors receiving in vitro fertilization (IVF) treatment were retrospectively enrolled to determine whether a history of endometriosis affects pregnancy outcomes. Patients with endometriosis had markedly fewer retrievable oocytes, a lower oocyte maturity rate, and decreased numbers of available and high-quality embryos (all p < 0.001) compared to those with tuber factors. The rates of clinical pregnancy and live birth in the endometriosis group were lower in the frozen-thawed embryo transfer cycles (p = 0.028 and p = 0.008, respectively), and a decreased cumulative live birth rate (CLBR) (p = 0.001) was observed. Logistic regression analysis revealed a negative association between endometriosis and CLBR (p = 0.002). The number of macrophages in the follicular fluid (FF) of patients with ovarian endometriosis was significantly higher than that of patients without ovarian endometriosis (p < 0.001). The levels of interleukin (IL)- 1α, IL-1ß, tumor necrosis factor-α, IL-6, IL-13, and IL-10 in FF were also elevated in the endometrioma group than in the control group (p < 0.05). These results indicate that endometriosis is negatively associated with CLBR in IVF, which may be caused by the follicular immune microenvironment that affects the development and fertilization of oocytes.

Bi-allelic mutations in MOS cause female infertility characterized by preimplantation embryonic arrest.

STUDY QUESTION: Are mutations in MOS (MOS proto-oncogene, serine/threonine kinase) involved in early embryonic arrest in infertile women? SUMMARY ANSWER: We identified mutations in MOS that may cause human female infertility characterized by preimplantation embryonic arrest (PREMBA), and the effects of the mutations in human embryonic kidney 293T (HEK293T cells) and mouse oocytes provided evidence for a causal relation between MOS and female infertility. WHAT IS KNOWN ALREADY: MOS, an activator of mitogen-activated protein kinase, mediates germinal vesicle breakdown and metaphase II arrest. Female MOS knockout mice are viable but sterile. Thus, MOS seems to be an important part of the mammalian cell cycle mechanism that regulates female meiosis. STUDY DESIGN, SIZE, DURATION: Whole-exome sequencing, bioinformatics filtering analysis and genetic analysis were performed to identify two different biallelic mutations in MOS in two independent families. The infertile patients presenting with early embryonic arrest were recruited from October 2018 to June 2020. PARTICIPANTS/MATERIALS, SETTING, METHODS: The female patients diagnosed with primary infertility were recruited from the reproduction centres of local hospitals. Genomic DNA from the affected individuals, their family members and healthy controls was extracted from peripheral blood. We performed whole-exome sequencing in patients diagnosed with PREMBA. Functional effects of the mutations were investigated in HEK293T cells by western blotting and in mouse oocytes by microinjection and immunofluorescence. MAIN RESULTS AND THE ROLE OF CHANCE: We identified the homozygous missense mutation c.285C>A (p.(Asn95Lys)) and the compound heterozygous mutations c.467delG (p.(Gly156Alafs*18)) and c.956G>A (p.(Arg319His)) in MOS in two independent patients. The mutations c.285C>A (p.(Asn95Lys)) and c.467delG (p.(Gly156Alafs*18)) reduced the protein level of MOS, and all mutations reduced the ability of MOS to phosphorylate its downstream target, extracellular signal-regulated kinase1/2. In addition, the identified mutations reduced the capacity of exogenous human MOS to rescue the metaphase II exit phenotype, and the F-actin cytoskeleton of mouse oocytes was affected by the patient-derived mutations. LIMITATIONS, REASONS FOR CAUTION: Owing to the lack of in vivo data from patient oocytes, the exact molecular mechanism affected by MOS mutations and leading to PREMBA is still unknown and should be further investigated using knock-out or knock-in mice. WIDER IMPLICATIONS OF THE FINDINGS: We identified recessive mutations in MOS in two independent patients with the PREMBA phenotype. Our findings reveal the important role of MOS during human oocyte meiosis and embryonic development and suggest that mutations in MOS may be precise diagnostic markers for clinical genetic counselling. STUDY FUNDING/COMPETING INTEREST(S): This work was supported by the National Natural Science Foundation of China (81725006, 81822019, 81771581, 81971450, 81971382,82001538 and 82071642), the project supported by the Shanghai Municipal Science and Technology Major Project (2017SHZDZX01), the Project of the Shanghai Municipal Science and Technology Commission (19JC1411001), the Natural Science Foundation of Shanghai (19ZR1444500 and 21ZR1404800), the Shuguang Program of the Shanghai Education Development Foundation and the Shanghai Municipal Education Commission (18SG03), the Foundation of the Shanghai Health and Family Planning Commission (20154Y0162), the Capacity Building Planning Program for Shanghai Women and Children's Health Service and the collaborative innovation centre project construction for Shanghai Women and Children's Health. The authors have no conflicts of interest to declare. TRIAL REGISTRATION NUMBER: N/A.

Optimal Endometrial Preparation Protocols for Frozen-thawed Embryo Transfer Cycles by Maternal Age.

This retrospective cohort study aimed to explore the optimal endometrial preparation protocols among different maternal age groups. A total of 16,867 frozen-thawed embryo transfer (FET) cycles were categorized into three groups based on endometrial preparation protocols: Natural cycle (NC n = 3893), artificial cycles (AC, n = 11456) and AC with GnRH-a pretreatment (AC+GnRH-a, n = 1518). To account for repeat cycles, a generalized estimating equation (GEE) method was applied to examine the associations between cycle regimens and pregnancy outcomes. Subgroup analyses were conducted to evaluate the best preparation methods for different maternal age groups. Primary outcomes were live birth and early miscarriage rates. After completing GEE, in overall population, the live birth rate [(NC as reference; AC: adjusted odds ratio (aOR) = 0.837, 95% confidential interval (CI) 0.771-0.908; AC+GnRHa: aOR = 0.906, 95%CI 0.795-1.031)] in NC was significantly higher than that in AC, while comparable that in AC+GnRH-a. The early miscarriage rate (AC: aOR = 1.420, 95%CI 1.225-1.646; AC+GnRHa: aOR = 1.545, 95%CI 1.236-1.931) was significantly lower in NC compared to either AC group. Subgroup analysis showed that in younger women, the incidences of live birth (AC: aOR = 0.900, 95%CI 0.804-1.007; AC+GnRHa: aOR = 1.091, 95%CI 0.904-1.317) were equivalent between groups, with a slightly higher in AC+GnRH-a. Early miscarriage rate (AC: aOR = 1.462, 95%CI 1.165-1.835; AC+GnRHa: aOR = 1.137, 95%CI 0.948-1886) was only significantly lower in NC compared to that in AC. In older women, the live birth rate (AC: aOR = 0.815, 95%CI 0.722-0.920; AC+GnRHa: aOR = 0.759, 95%CI 0.627-0.919) was significantly higher, and early miscarriage rate (AC: aOR = 1.353, 95%CI 1.118-1.638; AC+GnRHa: aOR = 1.704, 95%CI 1.273-2.280) was significantly lower in NC compared to either AC group. Our study demonstrated that NC is associated with lower early miscarriage late in overall IVF population. There is a mild favor of AC+GnRH-a in younger women, while the priority of NC is remarkable in older women. Maternal age should be a considerable factor when determining endometrial preparation method for FET.

Diagnosis of chronic endometritis: How many CD138+ cells/HPF in endometrial stroma affect pregnancy outcome of infertile women?

PROBLEM: The definition of chronic endometritis (CE) differs among studies, and currently, there is no accepted consensus. This study aimed to establish the minimum number of immunohistochemical analysis of CD138+ plasma cells to identify a clinically relevant CE. METHOD OF STUDY: We performed a retrospective study on 716 infertile patients who never did CE analysis and respective antibiotic treatment before. Samples were obtained by endometrial scratching in the mid-luteal phase before IVF-ET treatment. The number and distribution of CD138+ cells were analyzed by immunohistochemistry. Thirty high-power fields (HPF) were evaluated for each sample. Patients were classified in 2 main groups: (a) CD138low (<5 CD138+ cells in all HPFs), (b) CD138high (≥5 CD138+ cells in at least one HPF). Pregnancy outcome was compared among the groups. RESULTS: In the CD138high group, ß-hCG positive rate, clinical pregnancy rate and live birth rate were significantly decreased (P = .04, P = .01, P = .04, respectively). Also after adjusting for patient age, body mass index (BMI), and clinical characteristics, the ß-hCG positive rate (P = .05), clinical pregnancy rate (P = .01) and live birth rate (P = .02) were significantly lower in the CD138high than those in the CD138low group. Within the CD138low group, these parameters were not significantly different between patients without any plasma cells and patients with up to 4 plasma cells/HPF. CONCLUSION: We conclude that immunohistochemical analysis of CD138+ cells is a reliable method to detect CE which can be identified by the presence of ≥5 plasma cells in at least one out of 30 HPF.

[Use of recombinant human luteinizing hormone for ovulation stimulation in in vitro fertilization-embryo transfer].

OBJECTIVE: To evaluate application of recombinant human luteinizing hormone (r-hLH) used in ovarian stimulation of assisted reproductive technique and impact on outcome of pregnancy. METHODS: From Apr. to Jul. 2009, 123 patients with low LH level (< 1 U/L) at day 3 of menstruation and down-regulation of pituitary function undergoing in vitro fertilization-embryo transfer (IVF-ET) in Reproductive Medical Center, Provincial Hospital Affiliated to Shandong University were enrolled in this study, whom were classified into 66 cases treated by r-hLH in r-hLH group and 57 cases without r-hLH treatment in non-r-hLH group. In the mean time, 145 patients with normal level of serum LH (1 - 2 U/L) not given by r-hLH treatment and undergoing IVF-ET were matched as control group. Total amount of gonadotropin, estradiol levels and LH levels on the administration of human chorionic gonadotropin (hCG), number of oocytes retrieved, number of 2PN zygotes, rate of high quality embryos, the rates of implantation and clinical pregnancy were compared among these three groups. RESULTS: The level of serum LH on the day of hCG administration were (1.59 ± 0.77) U/L in r-hLH group, (0.54 ± 0.25) U/L in non-r-hLH group and (2.39 ± 1.01) U/L in control group, which reached statistical difference between every two groups (P < 0.05). The rates of high quality embryo were 59.36% in r-hLH group, 57.79% in non-r-hLH group, which were significantly lower than 65.94% in control group, respectively (P < 0.05). The rates of 2PN were 67.62% in r-hLH group and 68.32% in control group, which were significantly higher than 62.84% in non-r-hLH group, respectively (P < 0.05). The rates of implantation of 29.77% in r-hLH group were significantly higher than 18.26% in non-r-hLH group (P < 0.05). The total amount of gonadotropin, estradiol level on the day of hCG administration, the number of oocytes retrieved, and clinical pregnancy rate were not significantly different among those three groups (P > 0.05). CONCLUSION: The administration of recombinant human luteinizing hormone in patients who are profoundly suppressed after down-regulation with long protocol can get more quality embryos, the higher rates of 2PN and implantation.