Four years of IVF/ICSI experience in Kampala (Uganda)

⁴Correspondence address. Zonlaan 49, 1700 Dilbeek, Belgium. E-mail: peterplatteau{at}telenet.be We all know that starting and running an ART clinic is not soeasy as some people might perceive from outside. Doing the samething in the middle of Africa is even more challenging as someevidences in the Western world are not so obvious in this partof the world. We started our clinic in Kampala in 2004. Theclinic was a converted apartment from a four flat building.In the beginning, we had difficulties with importing drugs,culture media and consumables; we had the feeling everybodywas against us. We overcame multiple power failures, night intrudersand a 20% masturbation failure, but once the first IVF/ICSIbabies were born, people started to believe in the project.At present, 250 IVF/ICSI cycles a year are done in batches,we have a successful embryo freezing programme, offer IUI/ICSIfor sero discordant HIV couples and have the first babies afterIVF, ICSI, testicular biopsy, embryo freezing, oocyte donationand surrogacy in Central Africa. The results are comparableto the ones in the Western world.     

体外受精-胚胎移植412个周期分析

IVF-ET(in vitro fertilization and embryo transfer)的临床妊娠率受多种因素的影响:妇女年龄,子宫内膜厚度,促性腺激素(Gonadtropin,Gn)的用量,移植胚胎数以及胚胎质量。回顾性分析本生殖中心2000年9月到2002年7月的IVF/ICSI(intra cytoplasmic sperm injection)共412个周期。并将所有周期按照年龄分成3组,随年龄的增长3组的妊娠率分别是42.31％、33.70％、28％;子宫内膜的厚度与临床妊娠率无显著性相关;促性腺激素的用量与临床妊娠率呈负相关(P<0.05);移植胚胎数及优质胚胎数与临床妊娠率呈显著性正相关(P<0.05)。     

A cytogenetic study of couples with recurrent spontaneous abortions and infertile patients with recurrent IVF/ICSI failure

PURPOSE:

This study was conducted to determine the frequency and contribution of chromosomal abnormalities in miscarriages and in couples with recurrent in vitro fertilization/intra cytoplasmic sperm injection (IVF/ICSI) failure.

MATERIALS and METHODS:

A total of 221 individuals; 79 with three or more recurrent spontaneous abortions and 142 with at least three IVF/ICSI failures. Chromosomal analysis from peripheral blood lymphocytes was performed according to standard cytogenetic methods using G-banding technique.

RESULTS:

Abnormal karyotype was found in 21 (9.50%) individuals. Of these 21 subjects, 4 (19.04%) exhibited sex chromosomal abnormalities and 17 (80.96%) had autosomal abnormalities. Male partners had significantly higher chromosomal abnormalities (5.88%) than of females (3.61%). These abnormalities were also higher in patients with recurrent spontaneous abortions than with IVF/ICSI failure (P < 0.05).

CONCLUSIONS:

These data may be indicative that chromosomal abnormalities are involved more in spontaneous abortions than in recurrent IVF/ICSI failure. Cytogenetic analysis could be valuable for these couples when clinical data fail to clarify the cause.     

Blood metal/metalloid concentration of male subjects undergoing IVF/ICSI treatment outcomes: A prospective cohort study

BackgroundPrevious epidemiology studies reported that heavy metal/metalloid exposure is associated with the impairment of semen quality. However, it is still not clear whether the in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) treatment outcome will be affected after the heavy metal/metalloid exposure of the male partners.MethodsA prospective cohort study with a 2-year followed-up was conducted in a tertiary IVF center. A total of 111 couples undergoing IVF/ICSI treatment were initially recruited from November 2015 to November 2016. Male blood concentrations of heavy metal/metalloid including Ca, Cr, Mn, Fe, Ni, Cu, Zn, As, Se, Mo, Cd, Hg, and Pb were measured by inductively coupled plasma mass spectrometry, and the lab and pregnancy outcome data were followed up. The associations between male blood heavy metal/metalloid concentration and the clinical outcomes were analyzed by Poisson regression analysis.ResultsOur results showed that none of the heavy metal/metalloid of male partners we investigated are significantly associated with the oocyte fertilization and good embryo (P ≥ 0.05); however, antral follicle count (AFC) was a protective factor for the oocyte fertilization (RR: 1.07, 95 % CI: 1.04–1.10). The blood Fe concentration of the male partner was positively associated (P < 0.05) with pregnancy in the first fresh cycle (RR:170.93, 95 % CI: 4.13–7082.04), cumulative pregnancy (RR: 23.61, 95 % CI: 3.25–171.64) and cumulative live birth (RR: 36.42, 95 % CI: 1.21–1092.54). In the first frozen embryo cycles, pregnancy was significantly associated (P < 0.05) with the blood Mn (RR: 0.01, 95 % CI:0.00–0.11) and Se concentration (RR: 0.01, 95 % CI:8.25 E-5–0.47) and female age (RR: 0.86, 95 % CI:0.75–0.99); live birth was significantly associated (P < 0.05) with the blood Mn concentration (RR: 0.00, 95 % CI: 1.14E-7–0.51).ConclusionsOur results suggested that the higher male blood Fe concentration was positively associated with pregnancy in the fresh embryo transfer cycle, cumulative pregnancy, and cumulative live birth, whereas the higher male blood Mn and Se concentration were associated with lower chance of pregnancy and live birth in the frozen embryo transfer cycle. However, the underline mechanism of this finding still needs further investigation.     

Attitudes Towards the Donation of Human Embryos for Stem Cell Research Among Chinese IVF Patients and Students

Bioethical debates on the use of human embryos and oocytes for stem cell research have often been criticized for the lack of empirical insights into the perceptions and experiences of the women and couples who are asked to donate these tissues in the IVF clinic. Empirical studies that have investigated the attitudes of IVF patients and citizens on the (potential) donation of their embryos and oocytes have been scarce and have focused predominantly on the situation in Europe and Australia. This article examines the viewpoints on the donation of embryos for stem cell research among IVF patients and students in China. Research into the perceptions of patients is based on in-depth interviews with IVF patients and IVF clinicians. Research into the attitudes of students is based on a quantitative survey study (n=427). The empirical findings in this paper indicate that perceptions of the donation of human embryos for stem cell research in China are far more diverse and complex than has commonly been suggested. Claims that ethical concerns regarding the donation and use of embryos and oocytes for stem cell research are typical for Western societies but absent in China cannot be upheld. The article shows that research into the situated perceptions and cultural specificities of human tissue donation can play a crucial role in the deconstruction of politicized bioethical argumentation and the (often ill-informed) assumptions about “others” that underlie socio-ethical debates on the moral dilemmas of technology developments in the life sciences.     

促排卵过程中添加重组人生长激素对卵巢储备功能低下患者IVF-ET结局的影响及妊娠结局的影响因素探讨

摘要 目的：探讨促排卵过程中添加重组人生长激素（r-hGH）对卵巢储备功能低下（DOR）患者体外授精-胚胎移植（IVF-ET）结局的影响,并分析妊娠结局的影响因素。方法：选择2021年1月至2022年1月徐州市中心医院拟接受IVF-ET治疗的DOR患者60例、徐州医科大学附属沭阳医院拟接受IVF-ET治疗的DOR患者36例,共计96例,采用随机数字表法分为两组,每组48例,对照组予以常规促排卵治疗,观察组促排卵过程中添加r-hGH治疗,比较两组促排卵、体外受精、胚胎移植以及妊娠相关指标。此外,根据妊娠结局将所有患者分成妊娠成功组和妊娠失败组,采用多因素Logistic回归分析IVF-ET结局的影响因素。结果：观察组胚胎种植率、临床妊娠率高于对照组（P＜0.05）;观察组与对照组注射重组人促卵泡激素（Gn）天数、Gn用量、人绒毛膜促性腺激素（HCG）日雌二醇（E2）水平、HCG日黄体生成素（LH）水平、HCG日子宫内膜厚度、获卵数、成熟卵数、受精率、卵裂率、可移植胚胎数、优质胚胎数、移植胚胎数、早期流产率比较差异无统计学意义（P＞0.05）。单因素分析结果显示：妊娠失败组女方平均年龄大于妊娠成功组,基础卵泡刺激素（FSH）水平、IVF周期≥2个的患者比例高于妊娠成功组,促排卵应用了r-hGH的患者比例低于妊娠成功组,HCG日子宫内膜厚度小于妊娠成功组,基础窦卵泡数（AFC）个数、成熟卵数、优质胚胎数少于妊娠成功组,差异均有统计学意义（P＜0.05）。多因素Logistic回归分析结果显示：女方年龄≥35岁、基础FSH≥14.40 IU/L、HCG日子宫内膜厚度＜10 mm、基础AFC＜5.5个是DOR患者IVF-ET妊娠失败的危险因素,而促排卵应用r-hGH治疗是保护因素（P＜0.05）。结论：促排卵过程中增加r-hGH治疗可提高DOR患者IVF-ET胚胎种植率、临床妊娠率。女方年龄、基础FSH和AFC、HCG日子宫内膜厚度、r-hGH治疗均与IVF-ET妊娠结局有关。     

Factors Associated with Failed Treatment: an Analysis of 121,744 Women Embarking on Their First IVF Cycles

Background

In-vitro fertilization (IVF) is the treatment of choice for unresolved infertility. It comprises a number of key steps, each of which has to be negotiated before the next is attempted, but the factors which are associated with failure at each stage have not been reported.

Methods and Findings

We analyzed anonymised national data on women undergoing their first fresh autologous IVF and intracytoplasmic sperm injection (ICSI) cycle in the United Kingdom between 2000 and 2007 to predict factors associated with overall lack of livebirth as well as the chance of non-progress at different stages of an IVF cycle. A total of 121,744 women were included in this analysis. Multivariable models underlined the importance of increased female age and duration of infertility, lack of previous pregnancy, and a diagnosis of tubal or male factor infertility in predicting the risk of not having a live birth in an IVF treatment. At each stage, a woman’s chance of proceeding to the next stage of IVF treatment is affected by increased age and duration of infertility. The intention to use intra-cytoplasmic sperm injection (ICSI) is associated with a decreased risk of treatment failure in women starting an IVF cycle (RR 0.93, 99% CI 0.92, 0.94) but this association is reversed at a later stage once fertilisation has been confirmed (RR=1.01, 99%CI 1.00, 1.03).

Conclusions

Female age is a key predictor of failure to have a livebirth following IVF as well as the risk of poor performance at each stage of treatment. While increased duration of infertility is also associated with worse outcomes at every stage, its impact appears to be less influential. Women embarking on ICSI treatment for male factor infertility have a lower chance of treatment failure but this does not appear to be due to increased chances of implantation of ICSI embryos.     

Clinical implications of sperm DNA damage in IVF and ICSI: updated systematic review and meta-analysis

The clinical effect of sperm DNA damage in assisted reproduction has been a controversial topic during recent decades, leading to a variety of clinical practice recommendations. While the latest European Society of Human Reproduction and Embryology (ESHRE) position report concluded that DNA damage negatively affects assisted reproduction outcomes, the Practice Committee of the American Society for Reproductive Medicine (ASRM) does not recommend the routine testing of DNA damage for in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI). Herein, our aim was to perform a systematic review and meta-analysis of studies investigating whether sperm DNA damage affects clinical outcomes in IVF and ICSI, in order to contribute objectively to a consistent clinical recommendation. A comprehensive systematic search was conducted according to PRISMA guidelines from the earliest available online indexing year until March 2020, using the MEDLINE-PubMed and EMBASE databases. We included studies analysing IVF and/or ICSI treatments performed in infertile couples in which sperm DNA damage was well defined and assessed. Studies also had to include information about pregnancy, implantation or live birth rates as primary outcomes. The NHLBI-NIH quality assessment tool was used to assess the quality of each study. Meta-analyses were conducted using the Mantel–Haenszel method with random-effects models to evaluate the Risk Ratio (RR) between high-DNA-damage and control groups, taking into account the 95% confidence intervals. Heterogeneity among studies was evaluated using the I² statistic. We also conducted sensitivity analyses and post-hoc subgroup analyses according to different DNA fragmentation assessment techniques. We identified 78 articles that met our inclusion and quality criteria and were included in the qualitative analysis, representing a total of 25639 IVF/ICSI cycles. Of these, 32 articles had sufficient data to be included in the meta-analysis, comprising 12380 IVF/ICSI cycles. Meta-analysis revealed that, considering IVF and ICSI results together, implantation rate (RR = 0.74; 95% CI = 0.61–0.91; I² = 69) and pregnancy rate (RR = 0.83; 0.73–0.94; I² = 58) are negatively influenced by sperm DNA damage, although after adjustment for publication bias the relationship for pregnancy rate was no longer significant. The results showed a non-significant but detrimental tendency (RR = 0.78; 0.58–1.06; I² = 72) on live birth rate. Meta-analysis also showed that IVF outcomes are negatively influenced by sperm DNA damage, with a statistically significant impact on implantation (RR = 0.68; 0.52–0.89; I² = 50) and pregnancy rates (RR = 0.72; 0.55–0.95; I² = 72), although the latter was no longer significant after correction for publication bias. While it did not quite meet our threshold for significance, a negative trend was also observed for live birth rate (RR = 0.48; 0.22–1.02; I² = 79). In the case of ICSI, non-significant trends were observed for implantation (RR = 0.79; 0.60–1.04; I² = 72) or pregnancy rates (RR = 0.89; 0.78–1.02; I² = 44), and live birth rate (RR = 0.92; 0.67–1.27; I² = 70). The current review provides the largest evidence to date supporting a negative association between sperm DNA damage and conventional IVF treatments, significantly reducing implantation and pregnancy rates. The routine use of sperm DNA testing is therefore justified, since it may help improve the outcomes of IVF treatments and/or allow a given couple to be advised on the most suitable treatment. Further well-designed controlled studies on a larger number of patients are required to allow us to reach more precise conclusions, especially in the case of ICSI treatments.     

In vitro production of llama (Lama glama) embryos by IVF and ICSI with fresh semen

The interest for South American camelids has increased in the last years. The aim of the present research was to compare the in vitro production of Lama glama embryos using two techniques: in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). For IVF technique, we compared the effect of adding or not, heparin, penicillamine and hypotaurine as sperm capacitating agents. In the oocyte group subjected to ICSI, activation with or without, ionomycin and 6-dimethylaminopurine (6-DMAP) was assessed. Semen samples were obtained by electroejaculation and incubated at 38 degrees C in a 25% (v/v) collagenase solution. The cleavage and embryo development rates were compared between the different experimental groups. Only the number of cleaved oocytes was less when ICSI with no activation was used (p<0.05).     

黄体酮对子宫内膜异位症在位内膜预处理与IVF治疗结局关系

摘要 目的：研究黄体酮对子宫内膜异位症在位内膜预处理改善IVF结局。方法：男性因素并卵巢子宫内膜异位囊肿168例,分两组：（1）实验组：IVF前自然周期月经第12天地屈孕酮30 mg/日、14天,3个月;（2）对照组：IVF前无干预。IVF前测血CA125,黄体期长方案促排卵,排卵后一周（垂体降调日）再测CA125并取子宫内膜行ER、PR、HOXA-10mRNA检测。HCG日测子宫内膜厚度、形态、血流。比较临床资料及结局、症状疼痛评分。结果：实验组胚胎种植率、临床妊娠率高于对照组（P<0.05）;HCG日对照组子宫内膜厚于实验组。实验组子宫内膜A型血流比率高于对照组（P<0.05）。实验组A型子宫内膜比率高于对照组,但无明显差异（P>0.05）;IVF前两组CA125均高于参考值,但无明显差异（P>0.05）。垂体降调日复查CA125,实验组明显低于对照组,实验组治疗后低于治疗前;实验组子宫内膜ER、PR、HOXA-10 mRNA表达量高于对照组,实验组分泌期子宫内膜比率高于对照组（P<0.05）;治疗后两组各项症状疼痛评分均较治疗前改善,且实验组优于对照组（P<0.05）。结论：IVF治疗中合并卵巢子宫内膜异位囊肿用黄体酮预处理在位内膜,可以降低血CA125,有利于转变子宫内膜组织类型、减小子宫内膜厚度、增加子宫内膜血流、增加子宫内膜ER、PR、HOXA-10 mRNA表达,改善在位内膜容受性,缓解症状疼痛,提高临床妊娠率。     

The Influence of Polyvinylpyrrolidone on Freezing of Bovine IVF Blastocysts Following Biopsy

A study was conducted to develop a better freezing protocol for in vitro developed biopsied bovine blastocysts. Biopsied blastocysts were exposed to 1.8 M ethylene glycol (EG) + 0.05 M trehalose (T) and different concentration (5, 10, and 20%) of polyvinylpyrrolidone (PVP). Exposure to the solutions alone did not affect their in vitro development (Experiment 1). Experiments 2, 3, and 4 tested the viability of biopsied blastocysts cryopreserved in 1.8 M EG + different concentrations of T (0, 0.05, 0.1, and 0.3 M), 1.8 M EG + different concentrations of PVP (0, 5, 10, and 20%), and 1.8 M EG + 0.05 M T + different concentrations of PVP (0, 5, 10, and 20%), respectively. The proportion of biopsied blastocysts that reexpanded following cryopreservation in 1.8 M EG + 0.05 M T (38.5%) and 1.8 M EG + 0.1 M T (36.1%) was significantly (P < 0.05) higher than the proportion that reexpanded in 1.8 M EG + 0.3 M T (13.9%) (Experiment 2). The viability and the percentage of embryos that developed to >250 μm in diameter in the 5, 10, and 20% PVP groups (77.8 and 50.0%, 78.1 and 43.8%, 76.9 and 65.4%, respectively) were significantly higher than those that developed cryopreserved without PVP (55.1 and 20.7%) (Experiment 3). Optimum development of in vitro culture of frozen-thawed biopsied blastocysts was obtained using 1.8 M EG + 0.05 M T and 20% PVP. Analysis of blastocysts >250/μm in diameter showed that the number of ICM cells of biopsied blastocysts cryopreserved in 1.8 M EG + 0.05 M T with or without PVP was not different from the number of unfrozen biopsied blastocysts. These results indicate that PVP has some beneficial effect on freezing of biopsied bovine blastocysts.     

Microorganisms within Human Follicular Fluid: Effects on IVF

Our previous study reported microorganisms in human follicular fluid. The objective of this study was to test human follicular fluid for the presence of microorganisms and to correlate these findings with the in vitro fertilization (IVF) outcomes. In this study, 263 paired follicular fluids and vaginal swabs were collected from women undergoing IVF cycles, with various causes for infertility, and were cultured to detect microorganisms. The cause of infertility and the IVF outcomes for each woman were correlated with the microorganisms detected within follicular fluid collected at the time of trans-vaginal oocyte retrieval. Microorganisms isolated from follicular fluids were classified as: (1) ‘colonizers’ if microorganisms were detected within the follicular fluid, but not within the vaginal swab (at the time of oocyte retrieval); or (2) ‘contaminants’ if microorganisms detected in the vagina at the time of oocyte retrieval were also detected within the follicular fluid. The presence of Lactobacillus spp. in ovarian follicular fluids was associated with embryo maturation and transfer. This study revealed microorganisms in follicular fluid itself and that the presence of particular microorganisms has an adverse affect on IVF outcomes as seen by an overall decrease in embryo transfer rates and pregnancy rates in both fertile and infertile women, and live birth rates in women with idiopathic infertility. Follicular fluid microorganisms are a potential cause of adverse pregnancy outcomes in IVF in both infertile women and in fertile women with infertile male partners.     

改良透射电镜方法观察小鼠ICSI胚胎核仁超微结构

小鼠植入前胚胎的发育过程中,核仁经历从简单到复杂、从致密结构到网状结构的变化。对核仁超微结构的观察有助于揭示早期胚胎发育过程中核仁结构的动态变化及其特定阶段的功能。但由于核仁结构微小,数目较少,并且在胚胎中只处于卵裂球细胞核的内部,难以定位,因而给核仁的超微结构观察带来很大的困难。本实验探索了透射电镜观察小鼠植入前胚胎核仁的方法:先用琼脂对小鼠胚胎进行预包埋,在经过常规的透射电镜样品制备流程后,将整个胚胎先切成半薄切片;经过甲苯胺蓝染色后,选取含核仁结构的切片进行重包埋;最后再对回收来的半薄切片进行超薄切片,醋酸铀染色后上电镜观察;最终成功获得小鼠胚胎植入前发育不同时期核仁清晰的透射电镜图像。     

Human granulosa cells in vitro: influence of GnRH/GnRH-agonist on steroidogenesis and on IVF outcome

Steroidogenic activities of the granulosa cells (GCs) from 84 IVF trials were evaluated with respect to a set of ovarian stimulation regimens. Oestradiol (E2) synthesis of the GCs in vitro (obtained at oocyte retrieval) was compared to the maximal serum E2 levels of the same patients at induction of ovulation. Three stimulation regimens were employed: human post-menopausal gonadotrophin (hMG) alone; hMG accompanied by daily doses of a gonadotrophin releasing hormone agonist (GnRH-a); hMG preceded by a single depot application of the GnRH-a. Plots of E2 synthesis in vitro against serum E2 levels indicated that the GnRH-a directly inhibited E2 synthesis in the granulosa cells. This was confirmed in vitro by adding the agonist to the culture medium: both progesterone (P) and E2 syntheses were reduced in the presence of GnRH-a. Despite this drawback, the success of in vitro fertilization (IVF), as gauged by pregnancies achieved, was best for the group which received the GnRH-a as a single depot dose during the previous menstrual cycle, prior to the commence of stimulation. This success is attributed to the lower incidence of cancellations because of premature leuteinizing hormone (LH) surges which happen sometimes during ovarian stimulation. The implications of a direct influence of GnRH-a on E2 synthesis need to be further investigated.     

双重套式PCR对不同取样胚胎性别鉴定灵敏度测试

目的:测定双重套式PCR微量扩增体系的灵敏度,为附植前遗传学诊断 (PGD)、家畜胚胎性别鉴定等提供技术保障。方法:以桑椹胚卵裂球为模板进行扩增,建立昆明白小鼠特异的SRY ZFX双重套式PCR性别鉴定体系。按卵裂球数量的不同分为五组,对应的卵裂球个数分别为 1、2、3、4、5及以上。根据有效扩增结果得出双重套式PCR的灵敏度。结果:第 1组 (卵裂球个数为 1 ),有效检出率为 85 % ( 34 /40 ),污染率为 7 5 % ( 3 /40 ),漏检率为 7 .5 % ( 3 /40 )。随着卵裂球个数的增多,有效检出率逐渐升高,而污染率和漏检率则呈逐渐下降趋势。第 5组 (卵裂球达到 5及以上 ),有效检出率达到 1 0 0 %,漏检率为 0。对于第 2、3、4、5组而言,有效检出率及漏检率各组间并无显著差异 (P >0 . 0 5 );而第一组与其它各组间,有效检出率及漏检率存在显著差异 (P <0 .0 5 )。结论:双重套式PCR可以有效扩增基因组DNA量约为 1 2pg的模板,且扩增片段为单拷贝片段。