Statistical modelling of data from the embryo transfer process of In-Vitro Fertilization (IVF) treatments is motivated by the need to perform statistical inference for potential factors and to develop predictive models for these treatments. The biggest issue arising when modelling these treatments is that a number of embryos are transferred but unless all of the embryos get implanted or fail to implant then it is not possible to identify which of the embryos implanted. Little work has been done to address this partial observability of the outcome as it arises in this context. We adopt an Embryo-Uterus (EU) framework where a patient response has distinct uterine and embryo components. This framework is used to construct statistical models, expand them to allow for clustering effects and develop a package that will enable the fitting and prediction of these models in STATA. The capabilities of this package are demonstrated in two real datasets, aimed in investigating the effect of a new embryo prognostic variable and the effect of patient clustering in these treatments. In a simulation study EU models are shown to be capable of identifying a patient covariate either as a predictor of uterine receptivity or embryo viability. However a simulation case study shows that a considerable amount of information about the embryo covariate is lost due to the partial observability of the outcome. Further simulation work evaluating the performance of a number of proposed alternatives to the EU model shows that these alternatives are either biased or conservative. The partially observed cycles are finally considered as a missing data problem and two novel modelling approaches are developed which are able to handle the structure of these treatments. These novel models, based on multiple imputation and probability weighting, are compared to the EU model using simulation in terms of predictive accuracy and are found to have similar predictive accuracy to the EU model.

Philosophiae Doctor - PhD / Infertility is currently a serious problem that is escalating, not only in South Africa, but also worldwide. In Cape Town, a culturally diverse, urban community of approximately 1000 couples are referred to the Groote Schuur Hospital Infertility Clinic annually. Although infertility is primarily regarded as a medical condition, the treatments have emotional effects on infertile couples due to the recurring highs and lows that often accompany treatments. This study aimed to qualitatively explore and understand the emotional and psychological experiences of infertility and its treatments (specifically In Vitro Fertilisation and Embryo Transfer). Social constructionism is based on the premise that realities are not constructed in a vacuum but rather undergo a process whereby the subjective and inter-subjective experiences over time and through cultural processes come to be regarded as truths. These truths become internalised and function as lenses through which we see ourselves, compose and invent ourselves accordingly, making sense of what would otherwise have been chaotic and meaningless experiences. Additional aims were to examine women's experiences of infertility care whilst undergoing treatment and describe their experiences of coping with infertility and In Vitro Fertilisation and Embryo Transfer (IVF-ET). Semi-structured, in-depth individual interviews were conducted with 21 women presenting with primary infertility at a fertility clinic. This study utilised an ethnographic case study design. The results of the study suggested that women perceived themselves as not conforming to a dominant belief system that promotes motherhood as the most important role for women. The women described their 'failure' to fulfill socio-cultural expectations as emotionally turbulent. Some of the psychological responses to infertility included feelings of disappointment, shock, denial, devastation, anger, frustration, sadness, inadequacy, poor self-image and self-esteem. The women's personal accounts of their experiences of In Vitro Fertilisation and Embryo Transfer (IVF-ET) revealed that they found the treatment to be highly stressful, with emotional bouts of anxiety, nervousness, excitement and optimism. A psychological synopsis of infertility and IVF-ET is infertility and IVF-ET presents. The findings in this study suggest the need for the incorporation of 6 presented. This diagrammatic representation shows the intensity of the emotional rollercoaster that psychosocial intervention into infertility management. Greater attention to the psychological and emotional repercussions of infertility treatment could lead to a more personalised approach which, in turn, would optimise patient satisfaction and also prepare couples for the demands of the program by informing them about better ways of coping.

In this study, luteinized human granulosa cells (GC) obtained during in vitro fertilization procedures were used as a model system to evaluate the effects of ethanol (EtOH), a well‐known reproductive toxin, on epidermal growth factor (EGF) and gonadotropin‐stimulated steroidogenesis. Our results demonstrate that the basal progesterone (P4) and estradiol (E2) secretion by human GC in vitro was dependent on the ovarian stimulation protocol. EGF significantly enhanced P4, but not E2, secretion in human GC from clomiphene citrate (CC), human menopausal gonadotropin (hMG), and hMG/gonadotropin‐releasing hormone agonist (GnRH‐a)‐treated patients. The effects of EGF plus luteinizing hormone (LH) were additive in cells from the CC group, but less than additive in hMG and hMG/GnRH‐a groups. EtOH at 20 mM or more inhibited EGF stimulated P4 secretion in human GC from all three patient groups. EtOH inhibited P4 secretion stimulated by EGF and LH cotreatment in the CC and hMG/GnRH‐a groups, but not in human GC from the hMG‐treated patients. These results suggest that basal and EGF or LH‐stimulated P4 secretion by human GC, as well as the effects of EtOH, are profoundly influenced by the follicle's hormonal milieu.

This study evaluated the effect of ovum aging on the in vitro fertilizability of mouse ova. Over 1347 ova were evaluated. Serial trypsin digestion of in vitro and in vivo aged ova revealed an increase in zona digestion time (0.25% trypsin) beginning at 40 hr, which increased over a 40-hr period and resulted in the unfertilized zona becoming as "hard" as the fertilized embryo zona. In vitro fertilizability showed a rapid decrease as zona hardening occurred with loss of cortical granules as assessed by electron microscopy. These data suggest that the window of fertilizability is "closed" by a spontaneous zona reaction occurring at about 55 hr post-human chorionic gonadotropin with loss of cortical granules and zona hardening as manifested by increasing zona digestion time with 0.25% trypsin.

The effect of RU 486 at different concentrations (1, 5, 10, and 20 µg/ml) was studied on sperm-egg interaction in vitro in B6D2Ff mice. The in vitro fertilization rate of mouse ova decreased from 77.0% (control) to 50.0%, 28.7%, and 7.5% in the presence of RU 486 concentrations at 5, 10, and 20 µg/ml medium, respectively (p < 0.001). A concentration of 1 µg/ml did not affect the fertilization rate. A progesterone concentration at equal to or double the concentration RU 486 did not reverse the inhibitory effect of RU 486 on in vitro fertilization, which suggests a progesterone-independent mechanism. Exposure of spermatozoa (for 90 minutes) or ova (for 60 minutes) to RU 486 (20 µg/ml) followed by washing with RU 486-free medium before coincubation did not affect the fertilization rate. The presence or absence of cumulus cells did not change the inhibitory effect of RU 466 (20 µg/ml) on in vitro fertilization. RU 486 at 5 to 20 µg/ml medium was associated with perivitelline polyspermy in nonfertilized ova and enhanced perivitelline polyspermy in fertilized oval.

<p>The experience of infertility and its associated treatments, including in vitro fertilization (IVF), can have a profound impact on the emotional health and well-being of women desiring to become mothers. While researchers have measured the impact of infertility or described the experience and processes related to infertility and its treatment, what remained to be explored was a virtually pre-packaged collection of rich descriptions of the IVF experience as captured in women’s blogs. This discourse analysis sought to describe the blogging practices of women undergoing IVF for treatment of infertility, exploring both the content and function of the IVF blog discourse. Data were collected from the text of seven women’s blogs and resulted in four main functions of the discourse: creation of and connection to a community, emotional support, blogging as therapy, and creation of an IVF resource. Findings suggest that blogging can have a positive impact on the psychosocial consequences experienced by women in fertility treatment. Findings of this study also have methodological implications for researchers considering blogs as a data source in qualitative research.</p> / Master of Science in Nursing (MSN)

Thesis (PhD)--Stellenbosch University, 2012. / ENGLISH ABSTRACT: CHAPTER 1 In this chapter the aim is to outline the different chapters under section A. Against this background, we will conduct a literature review of relevant studies performed, and evaluate their comments regarding identifying embryo markers which can be utilized to improve overall ART outcome. We will evaluate the embryo marker sHLA-G in detail, using a prospective randomized study as well as a retrospective multi-centre study. The role of the morphology and genetic profile of an oocyte, zygote and embryo and subsequent blastocyst formation will be evaluated with the help of WGA/CGH. The work will then be summarized and conclusions will be made as well as possible suggestions for future directions will be indicated. In section B the methodology of the studies explaining the role of the candidate is illustrated. CHAPTER 2 In this chapter the impact of the oocyte/zygote and the embryo on implantation/pregnancy rate was discussed. The morphologic characteristics of the oocyte, the cumulus–oocyte-complex (COC), the zona pellucida, the perivitelline space, cytoplasm and meiotic spindle and the polar body and its appearance were discussed in detail. The morphologic characteristics of embryo fragmentation and its effect on embryo development, ploidy and blastocyst formation were also studied. Embryo markers to predict pregnancy outcome were researched based on the international literature. The pronuclear morphology and early cleavage were highlighted as non-invasive embryo markers to predict outcome. A non-invasive biochemical marker, soluble Human Leucocyte-Antigen-G (sHLA-G), that is expressed by developing embryos was researched. The value of blastocyst transfer and the improved ongoing pregnancy rate compared to cleavage stage embryos were highlighted based on a recent meta-analysis. A detailed discussion on sHLA-G as well as Array-CGH and the future of these tests followed. CHAPTER 3 In this chapter the aim was to compare pregnancy and implantation rates when embryos are selected based on a single Day 3 (D 3) morphology score vs. a GES score plus sHLA-G expression. This was a prospective randomized study (n=214) undergoing fresh ICSI cycles. Embryos were selected for transfer based on either Day 3 morphology score (Group A) or GES-scoring plus sHLA-G expression (Group B). The following results were reported: Clinical [35/107 (33%) vs. 52/107 (49%)] and ongoing pregnancy [20/107 (19%) vs. 52/107 (49%)] rates were significantly different between Group A and Group B (p<0.05). Implantation rates were not significantly different between Group A [52/353 (15%)] and Group B [73/417 (18%)] (p<0.05). The number of pregnancies lost during the first trimester was nearly 12 times higher in Group A [25/52 (48%)]. It was concluded that the miscarriage rate was significantly lower in Group B than Group A and the pregnancy results were superior when embryos were selected based on GES plus sHLA-G expression. CHAPTER 4 Several studies have reported an association between the presence of soluble human leukocyte antigen-G (sHLA-G) in human embryo culture supernatants (ES) with implantation and pregnancy outcome in vitro. However, the actual presence role during implantation and effect on implantation and pregnancy outcome are still controversial. A retrospective multi-centre study was performed on 2040 ICSI patients in six different centers. All embryos were individually cultured and a chemiluminescence enzyme-linked immunosorbent assay (ELISA) was used to detect the presence of sHLA-G in culture medium surrounding embryos. In all centers, a positive sHLA-G result was associated with an increase in odds of multiple clinical implantations (OR: 1.48, 95% CI: 1.07 to 2.05, p-value: 0.0170), and an increased odds of multiple on-going pregnancies (OR: 1.66, 95% CI: 1.10, 2.51, p-value: 0.0170). Data from this multi-centre study emphasize that sHLA-G expression is a valuable non-invasive embryo marker to assist in improving pregnancy outcome with the theoretical potential to reduce multiple pregnancies. A combination of sHLA-G expression and extended embryo culture to the blastocyst stage might provide future tools by which to select single embryos for transfer and reduce the risk of multiple gestational, without compromising their pregnancy rates. CHAPTER 5 In this chapter the ploidy status of first and second polar bodies and Day 3 blastomere, embryo morphology and biochemical (sHLA-G) characteristics were correlated with blastocyst development and subsequent pregnancy outcome. All oocytes/zygotes and embryos were individually cultured to the blastocyst stage. PB-I, PB-II and blastomeres underwent whole genome amplification (WGA) and comparative genome hybridization (CGH) and complete karyotyping. Each embryo‟s culture medium supernatant was collected and analyzed for sHLA-G expression on Day 2. The following results were reported: Fifty seven mature (MII) donor oocytes were obtained, 33/57 (57.9%) were aneuploid, 21/57 (36.8%) were euploid and 3/57 (5%) were “inconclusive”. No correlation was found between CGH status of PB-I, PB-II and the GES-score. Furthermore, no correlation was established between PB-I CGH results and blastocyst morphology grade. There was a significant correlation between PB-I CGH and blastomere CGH results. Euploid and aneuploid PB-I developed into 58% and 67% blastocysts, respectively. Kappa statistics (>0.7) revealed a positive correlation between the ploidy of PB-I, PB-II and the blastomeres. It was concluded that following ICSI and sequential genetic karyotyping of the oocyte/zygote and subsequent blastomeres, the majority of oocytes fertilized and subsequent zygotes developed into blastocysts, despite their ploidy status. We therefore conclude that blastocyst development is not associated with ploidy. CHAPTER 6 Identifying a developmentally competent embryo to transfer that has the highest probability to develop into a live baby has been an issue of debate and continues research. The aim of this chapter is to discuss the morphological, biochemical and genetic features of an embryo that has been shown to be predictive of implantation and pregnancy outcome in ART using most current evidence. A literature search was performed looking at the correlation between pronuclear morphology, early cleavage, cleavage stage embryos, blastocyst development, the presence of sHLA-G, CGH, embryo development and implantation/pregnancy rates in ART. Based on the available literature, a combination of observations could assist the scientist with embryo selection. The pronuclear stage morphology, the early embryo division, cleavage embryo stage and quality of the day 3 embryos provides limited guidance. However, choosing a blastocyst with a positive sHLA-G result on Day 5 is the optimal combination to make the final selection before embryo transfer or freezing. This non-invasive approach should improve pregnancy outcome and reduce multiple pregnancy rates. As far as the use of the more invasive technology such as aCGH is concerned, more research on pregnancy outcome is needed. CHAPTER 7 A combination of observations for embryo selection, starting with oocyte grading, pronuclear stage morphology, early zygote cleaving and cleavage-stage embryo morphology/quality on Day-3, however, ultimately using extended embryo culture and choosing a blastocyst on Day 5 with positive sHLA-G values available, will assist the scientist in making the final decision before selecting an embryo for transfer or cryopreservation. The use of aCGH (for chromosomal analysis) is invasive and is still considered experimental. Finally we conclude that despite all the above mentioned parameters to select an embryo for transfer that will develop into a live baby, more extensive research and international corroboration is needed in order to improve and standardize embryo selection criteria. / AFRIKAANSE OPSOMMING: HOOFSTUK 1 Die doel in hierdie hoofstuk is om die verskillende hoofstukke onder Afdeling A uiteen te sit. Daar word beplan om „n literatuur oorsig te doen van toepaslike studies rakend embriomerkers wat swangerskap-uitkoms in in vitro bevrugting kan verbeter. Verder sal die embriomerker sHLA-G deeglike bestudeer word met behulp van „n prospektiewe gerandomiseerde studie, asook „n retrospektiewe multisentrum studie. Die rol van embrio morfologie en die genetiese profiel van die ovum, sigoot asook die embrio en die daaropvolgende blastosist vorming sal geëvalueer word met behulp van WGA/CGH. Alle bevindings sal daarna opgesom word, gevolg deur „n sinvolle gevolgtrekking en laastens sal voorstelle gemaak word vir toekomstige navorsing op die gebied. In Afdeling B sal die metodiek van die studies verduidelik word, asook „n beskrywing gegee word van die kandidaat se rol gedurende die navorsings projekte in hierdie tesis. HOOFSTUK 2 In hierdie hoofstuk word die impak van die oösiet en die embrio op die inplanting/swangerskap-koers bespreek. Die morfologiese eienskappe van die oösiet, die kumulus-oösiet kompleks, die sona pellucida, die perivitelline spasie, sitoplasma en meiotiese spoel, die poolliggaam en die se voorkoms word breedvoerig bespreek. Die morfologiese eienskappe van die embrio, fragmentasie en die invloed daarvan op die embrio, ploïdie, en blastosistvorming word bespreek. Embriomerkers om swangerskapsuitkoms te voorspel, gebaseer op internasionale literatuur, is ook nagevors. Die pronukleêre morfologie en vroeë deling word as nie-indringende embriomerkers uitgelig om swangerskapsuitkoms te voorspel. „n Biochemiese, nie-indringende merker wat deur ontwikkelende embrios uitgedruk word, oplosbare menslike leukosiet antigeen-G (sHLA-G), word bespreek. Die waarde van blastosist oordrag en die verbeterde koers van voortgaande swangerskappe in vergelyking met verdelende embrios, is ook uitgelig, gebaseer op „n onlangse metanalise. „n Breedvoerige bespreking van sHLA-G asook “Array-CGH” en die toekoms van hierdie toetse word behandel. HOOFSTUK 3 Die doel van hierdie hoofstuk is om swangerskap en inplantingskoerse te vergelyk wanneer embrios geselekteer word op „n enkel Dag 3 (D 3) morfologie beoordeling, teenoor „n kumulatiewe GES-telling plus sHLA-G uitdrukking. Hierdie was „n prospektiewe ewekansige studie (n=214) waar pasiënte ICSI-siklusse ondergaan het. Embrios is geselekteer vir terugplasing gebaseer op óf Dag 3 morfologie telling (Groep A), óf „n kumulatiewe GES-telling plus sHLA-G uitdrukking (Groep B). Die volgende resultate is gerapporteer: kliniese swangerskappe [35/107 (33%) vs 52/107 (49%)] en voortgaande swangerskappe [20/107 (19%) vs. 52/107 (49%)] se sukses koerse is beduidend verskillend tussen Groep A en Groep B (p<0.05). Inplantingskoerse is nie beduidend verskillend tussen Groep A [52/353 (15%)] en Groep B [73/417 (18%)] (p<0.05) nie. Die aantal swangerskappe wat tot niet gegaan het tydens die eerste trimester was bykans 12 keer hoër in Groep A [25/52 (48%)]. Die slotsom was dat die miskraamsyfer beduidend laer in Groep B as in Groep A is en die swangerskap syfer betekenisvol beter was wanneer die selektering van embrios op GES plus sHLA-G gebaseer is. HOOFSTUK 4 Verskeie studies het „n assosiasie getoon tussen die teenwoordigheid van oplosbare menslike leukosiet antigeen-G (sHLA-G) in menslike embrio kultuur en swangerskaps uitkoms in vitro. „n Retrospektiewe studie is op 2040 ICSI pasiënte by 6 verskillende sentra gedoen om die effek van s-HLAG verder te bestudeer. Alle embrios is individueel gekweek om die teenwoordigheid van sHLA-G in „n kultuurmedium rondom die embrios te identifiseer. In alle sentra is „n positiewe sHLA-G uitslag met „n toename in die waarskynlikheid van veelvuldige inplantings geassosieer (OR: 1.48, 95% CI: 1.07 tot 2.05, p-waarde: 0.0170), asook „n toename in waarskynlikheid van meervoudige swangerskappe wat voortduur (OR: 1.66, 95% CI: 1.10, 2.51, p-waarde: 0.0170). Data uit die multisentriese studie beklemtoon dat sHLA-G uitdrukking „n waardevolle nie-indringende embriomerker is om by te dra tot die verbetering van swangerskapsuitkoms, asook die teoretiese potensiaal om meervoudige swangerskappe te verminder. „n Kombinasie van sHLA-G uitdrukking en verlengde embrio kultuur tot die blastosist stadium mag moontlik „n toekomstige hulpmiddel wees waardeur enkele embrios vir terugplasing geselekteer kan word. Daardeur kan die risiko van meervoudige swangerskappe beperk word sonder om die swangerskapkoerse in gevaar te stel. HOOFSTUK 5 In dié hoofstuk word die ploïdie status van die eerste en tweede poolliggaampies en Dag 3 blastomere, embrio morfologie en biochemiese (sHLA-G) eienskappe gekorrelleer met blastosist ontwikkelling en uiteindelike swangerskapsuitkoms. Alle oösiete/sigote en embrios is individueel tot die blastosist stadium gevolg. PB-I, PB-II en blastomere het “volledige kariotipering ondergaan deur gebruik te maak van die toets “comparative genome hybridization (CGH)”. Elke embrio se kultuurmedium supernatant is versamel en ontleed vir sHLA-G uitdrukking op Dag 2. Die volgende uitslae is gerapporteer: Sewe-en-vyftig mature (MII) donor oösiete is verkry; 33/57 (57.9%) is aneuploïd, 21/57 (36.8%) is euploïd en 3/57 (5%) is onbeslis. Geen verwantskap is gevind tussen CGH status van PB-I, PB-II en die GES-telling. Geen verwantskap is gevind tussen CGH status van sHLA-G. Verder was daar geen verwantskap gevind tussen PB-I CGH uitslae en blastosist morfologie graad nie. Daar was „n beduidende korrelasie tussen PB-I CGH en blastomeer CGH uitslae. Euploïde en aneuploïde PB-I het onderskeidelik in 58% en 67% blastosiste ontwikkel. Daar is „n positiewe verwantskap tussen die ploïdie van PB-I, PB-II en die blastomere aangetoon [Kappa (>0.7)]. Dit is afgelei dat na ICSI en sekwensiële genetiese kariotipering van die oösiet/sigoot en daaropvolgende blastomere, die meerderheid oösiete bevrug is en die daaropvolgende sigote ontwikkel het tot blastosiste, ongeag hul ploïdie status. Ons afleiding is dus dat blastosist ontwikkelling nie aan ploïdie verwant is nie. HOOFSTUK 6 In hierdie hoofstuk bespreek ons waarnemings wat betref seleksie kriteria om die beste embrios te kies vir terugplasing wat uiteindelik tot „n suksesvolle swangeskap sal lei. Morfologiese, biochemiese en genetiese faktore is ondersoek. „n Onderskeiding is gemaak tussen nie-indringende (mikroskopiese en biochemiese) en indringende (embrio biopsie, aCGH) tegnieke. 'n Kombinasie van nie-indringende observasies, wat insluit pronukliere mofologie, vroee sigoot verdeling en vroeë embrio morfologie/kwalitieit op Dag-3 het beperkte inligting verskaf wat betref swangerskapkans. Verlengde embrio kweking tot die blastosist stadium (Dag-5) plus „n positiewe sHLA-G resultaat gee egter veel meer voordelige inligting aan die embrioloog met die embrio seleksie proses, voor embrio terugplasing of bevriesing. Laasgenoemde inligting sal die swangerskap syfer bevoordeel en die meervoudige swangerskap kans verlaag. Wat die indringende tegniek (aCGH) betref, word veel meer data benodig rakend die potensiele voor- en nadele wat betref swangerskap uitkoms, voordat „n sinvolle gevolgtrekking gemaak kan word. HOOFSTUK 7 „n Volledige literatuur oorsig dui daarop dat alle beskikbare riglyne om embrios te kies vir terugplasing, ingespan moet word. In die studie is daar gekyk na „n kombinasie van hierdie voorstelle. Daar is begin met die morfologie van die pronukliere stadium, gevolg deur vroeë sigoot-verdeling, asook beoordeling van embrios se morfologie/kwaliteit op Dag-3 van ontwikkeling. Daar word voorgestel dat die keuse van „n blastosist op Dag 5, gekombineerd met „n positiewe oplosbare menslike leukosiet antigeen G (shla-G) die embrioloog van hulp kan wees om die beste embrio te kies vir terugplasing of bevriesing. Hierdie nie-indringende riglyn behoort swangerskap-uitkoms te verbeter asook meervoudige swangerskappe te verminder. Indringende tegnieke soos ACGH benodig verdere in diepte navorsing en data verkryging om die waarde van hierdie toets te kan beoordeel.

Thesis (PhD(Agric) (Animal Sciences))--University of Stellenbosch, 2005. / As most researchers have foreseen, and many breeders have hoped, the in vivo and in vitro production of livestock embryos and the birth of subsequent offspring never really replaced artificial insemination during the past 30 years. This was, to a large extent, due to very variable and unreliable numbers of embryos produced using these two methods. The present study was therefore undertaken to investigate certain aspects of assisted reproductive techniques (ART) to try and solve some of these difficulties. Problems addressed were the management of follicular development on the ovary by controlling the dominant follicle, and investigating alternative and more cost-effective culture media and conditions for embryo culture. A method to control the development of the dominant follicle in a cohort of follicles as well as the waves of follicular development in the ovaries of sheep, goats and cows with an estrogenic product was investigated. Estradiol cypionate (ECP) was used for this purpose, injected intramuscularly after the insertion of the progesterone or progestagen implant. ECP has a negative feedback effect on the secretion of pituitary FSH, and therefore follicular development. The animals of the three different species were randomly divided into two groups each, the ECP-group receiving the estradiol cypionate injection, and the control group receiving a saline injection. Five days after the ECP injection a program of follicular multi-stimulation with FSH hormone was initiated. The females of the different species were bred by either natural service (goats) or inseminated by laparoscopy (sheep) or trans-cervically (cows) to fertilize the ovulated ova. Embryos and unfertilized ova were collected surgically at the 8 to 16- cell stage 3 to 4 d after breeding in the sheep and goats, and trans-cervically in the cows. Significantly more CL formed, and a total number of ova and embryos, as well as transferable embryos, were collected from the ECP-group of sheep ewes and goat does compared to the control group that received no ECP (p<0.01). There was, however, no difference in the average number of unfertilized ova that were collected in the two sheep or goat groups. In the cows the number of CL counted, the total number of embryos and ova and of transferable embryos collected, were significantly greater (p<0.05) in the group that were injected with ECP compared to the group that received no ECP. The control group also had a significantly larger number of unfertilized ova than the ECP-group (p<0.05). It could therefore be concluded that more reliable numbers of embryos can be produced in vivo if the development of the dominant follicle as well as the subordinate follicles is controlled with estradiol cypionate. Since more than half a century ago, attempts have been made to culture cells and embryos outside the body (in vitro or ex vivo). This was done with different culture media and in various "incubators". Chapter 2 deals with two different culture media used: a standard TCM-199 culture medium and first trimester amniotic fluid (BAF) collected sterilely from pregnant cows after slaughter. Two different culture conditions were also investigated, the standard laboratory CO2 incubator versus culturing bovine embryos in the vagina of a goat doe. Two experiments were done: Firstly the permeability of different receptacles to CO2 gas was analyzed for possible culture in the vagina. Four-well plates and straws were used to incubate TCM-199 and BAF for a period of 120 h in the presence or absence of 5% CO2 gas. The pH values were measured every 24 h and recorded. In the second experiment pre-compacted morula stage bovine embryos were incubated in the above culture media in sealed 0.25 mL straws in a standard laboratory incubator and in the vagina of a goat doe. Evaluation was done on (1) stage of development and (2) number of blastomeres after 96 h of culture. In experiment one it was shown that the CO2 gas diffused out of the 4-well plate as well as the straws in the absence of CO2 gas, while in the presence of CO2, the pH of both media stabilized between 7.3 and 7.5. This meant that the semen straws were permeable to CO2 gas and could therefore be used as receptacles for culturing early stage bovine embryos. In the second experiment no statistical differences (p>0.05) were found in the number of Grade 1 pre-compacted bovine embryos that developed to the blastocyst stage, or the hatched blastocyst stage, neither for the culture medium used, or the method of culturing in the two incubators. Neither was there any difference (p>0.05) in the number of blastomeres that developed at the blastocyst stage between the two types of incubators used. Embryos tended to develop more blastomeres when cultured in BAF than when cultured in TCM-199 in both the standard laboratory incubator and when using the vagina of a goat doe as an incubator (p<0.05). After the collection of in vivo produced livestock embryos, they are evaluated under high magnification (minimum of 80X) with the aid of an inverted or stereo microscope. The Grade 1 embryos will give the best conception results when transferred to synchronized recipient female animals, while the Grade 3 embryos will give the worst results. The aim of the next experiment was to culture all three quality grades of in vivo produced pre-compacted morula-stage embryos of sheep, goats and cows in two different culture media and then compare the development of the embryos by evaluating the number of embryos reaching the hatched blastocyst stage. The results have shown that there were no significant differences between the development of the Grade 1 and the Grade 2 embryos from any of the three species when either cultured in TCM- 199 or heat inactivated early pregnancy-stage (<60 d) bovine amniotic fluid (BAF) were used as culture media. Significantly more in vivo produced Grade 3 pre-compacted morula-stage sheep, goat and cow embryos, however, developed to the hatched blastocyst stage when cultured in BAF with 10% FBS and antibiotics, compared to culture in TCM-199 with 10% FBS and antibiotics (p<0.05). The effect of co-culture on the survival of caprine embryos post transfer to a synchronized recipient female goat was also assessed. A total of 120 Kashmir embryos at the blastocyst stage were divided into three groups after thawing and reconstitution in four steps in glycerol and sucrose medium. The first group of embryos (G1, n=40) was individually transferred semi laparoscopically in D-PBS with 10% FBS and antibiotics to the ipsilateral horn of the CL over a period of 3 d. The second group of caprine blastocysts (G2, n=40) was similarly transferred in TCM-199 with FBS and antibiotics. The third group of frozen-thawed caprine blastocyst-stage embryos (G3, n=40) were first co-cultured for ~24 h in TCM-199 with serum and antibiotics in groups of up to five embryos inside a ~50 mm length of a semen straw in a cylindrical sponge in the anterior part of the vagina of a goat doe in her luteal phase. After the culture period these embryos were transferred in a similar way in TCM-199 without the co-culture as in G1 and G2. Ultrasound scanning showed that significantly more of the blastocyst embryos that were cocultured in the vagina (G3) before transfer developed to a pregnancy compared with the embryos transferred in D-PBS (G1). The co-culture Group 3 blastocyst-stage caprine embryos produced significantly more offspring than the non-cultured embryos transferred in both D-PBS (G1) and TCM-199 (G2) (p<0.05). The maturation of bovine oocytes to allow the oocyte to resume meiosis, is the first step in in vitro fertilization to produce IVMFC embryos. The composition of the maturation medium plays an important role in the success achieved with maturation. An investigation was therefore launched to evaluate the maturation ability of first trimester bovine amniotic fluid (BAF) to mature prophase I oocytes collected from abattoir ovaries to metaphase II oocytes, compared to a standard maturation medium such as TCM-199. In the first experiment three groups of ~100 oocytes each were matured in TCM-199 with estrus cow serum (ECS). The first group of oocytes was matured in a 50 μL micro-drop in an incubator, while the other two groups were matured in semen straws, one group in an incubator and the other group in the vagina of a goat doe in di-estrus. Six further groups of ~100 oocytes each, with BAF as maturation medium, three groups with ECS and three without ECS, were matured in the same receptacles and under the same conditions as with the TCM-199. No significant differences in number of oocytes reaching the metaphase II stage could be found for any of the nine treatment groups. In the ...

<p>The chromatin integrity in sperm cells is vital for successful pregnancy. In this</p><p>study DNA-damage was evaluated in sperm cells from 50 men attending In Vitro Fertilization</p><p>(IVF) or Intra Cytoplasmic Sperm Injection (ICSI) treatment. Male semen samples were</p><p>purified with a two-shift gradient before the sperm cells were treated with the Halosperm® Test</p><p>Kit and evaluated for DNA-damage. The samples were divided in two groups according to DNAFragmentation</p><p>Index (DFI) of 30 % and the results correlated with fertilization, cleavage and</p><p>pregnancy rate. Men with DFI ≥ 30 % had a higher fertilization and pregnancy rate and a lower</p><p>cleavage rate compared to men with DFI ≤ 30 %. The conclusions were that fertilization in vitro</p><p>may be independent of the degree of DNA-damage, the embryonic development could be</p><p>seriously disrupted by damaged sperm cells, and the pregnancy rate showed no correlation to a</p><p>DFI threshold of 30 %.</p>

Infertilitet är en utbredd problematik som drabbar många par världen över. Olika behandlingsformer finns att tillgå för att hjälpa infertila par att uppnå en graviditet. IVF-behandling är en vanlig behandlingsform. Många studier visar att IVF-behandling påverkar kvinnan mer än mannen psykiskt. Syftet med denna studie var att beskriva hur kvinnor upplever processen att genomgå IVF-behandling. Metod: En litteraturstudie gjordes på tio kvalitativa artiklar för att besvara syftet. En innehållsanalys gjordes på dessa artiklar, där meningsbärande enheter identifierades i artiklarnas resultat. Dessa enheter kondenserades, grupperades och formade sub-kategorier och teman. Livsvärldsperspektivet användes som en teoretisk referensram. Resultatet av denna litteraturstudie blev sju teman; Tro på behandlingen; Okunskap; Påverkan på vardagen; Bemötande i relationer; Negativ hantering; Rädsla, samt Väntan. Slutsatser som drogs av denna litteraturstudie var att kvinnornas upplevelser var övervägande negativt präglade, men en tro på behandlingen var ändå en central upplevelse. Det är viktigt för berörda kvinnors omgivning att inneha ett livsvärldsperspektiv inför kvinnornas upplevelser av behandlingen. En ökad förståelse från kvinnornas omgivning skulle kunna ge kvinnorna bättre upplevelser av att genomgå behandlingen. / Infertility is a widespread problem that affects many couples worldwide. Various treatments are available to help infertile couples achieve conception, and IVF is a common form of treatment. The psychological impact of the IVF-treatment affects the woman more than the man. The aim of this study was to describe how women experience the process of going through IVF treatment. Method: A literature review was done on ten qualitative articles to answer the aim of the study. A content analysis was done on these articles, and meaningful units were identified in the articles‟ results. These units were condensed, grouped and then formed into sub-categories and themes. A Life-World perspective was used as a theoretical framework. The result of this study was seven themes: Faith in the treatment; Ignorance: The impact on daily life; Responding in relationships, Negative handling, Fear, and Wait. Conclusions of this study were that women's experiences were predominantly negatively characterized, but a faith in the treatment was still a central experience. It is important for the women‟s surroundings to hold a Life-World perspective to women's experiences of the treatment. A greater understanding from the women's surroundings may improve the women‟s experiences of going through the treatment.