Projects / Programmes
The effect of non-apoptotic sperm selection using MACS method on the epigenetic status of spermatozoa and on the outcome of in vitro fertilization procedure
| Code |
Science |
Field |
Subfield |
| 3.05.00 |
Medical sciences |
Human reproduction |
|
| Code |
Science |
Field |
| B007 |
Biomedical sciences |
Medicine (human and vertebrates) |
| Code |
Science |
Field |
| 3.02 |
Medical and Health Sciences |
Clinical medicine |
infertility, sperm selection, apoptosis, epigenetics, assisted reproduction
Researchers (1)
| no. |
Code |
Name and surname |
Research area |
Role |
Period |
No. of publicationsNo. of publications |
| 1. |
33917 |
PhD Martin Štimpfel |
Human reproduction |
Head |
2017 - 2019 |
85 |
Organisations (1)
Abstract
This study will be focused on two important aims. First aim is to identify the importance of new method - this is the selection of non-apoptotic spermatozoa for in vitro fertilization procedure using MACS, on the development and implantation of embryos and on the pregnancy outcome. The second aim is to determine how unselected and MACS-selected spermatozoa population differ on epigenetic level. We will perform a prospective study in couples included in an infertility treatment programme at our IVF Centre with male factor of infertility, more precisely, in patients with teratozoospermia. In each couple one half of the mature oocytes will be fertilized with ICSI using spermatozoa prepared with routine sperm preparation method, and the second half will be fertilized with ICSI using spermatozoa selected using MACS procedure (MACS-ICSI). For the MACS procedure the commercially available kit (MACS® ART Annexin V System) will be used. Embryos will be cultured individually in a single-step culture medium (in a time lapse system) to enable optimal monitoring of their development. After five days of culture, one the most optimal embryo (blastocyst) will be transferred into the uterus. The surplus blastocysts will be cryopreserved and cryostored for later use. The quality of blastocysts (development, morphology of embryoblast and trophoblast) will be precisely evaluated to enable the comparison between blastocyst regarding to the procedure used for spermatozoa selection.
To evaluate the epigenetic status of the spermatozoa, they will be sampled in two groups from each included patient. In the first group there will be samples of spermatozoa prepared only with the routine sperm preparation method, and in the second group of samples spermatozoa selected using the MACS procedure. Using this approach we will be able to evaluate whether the epigenetic status of spermatozoa varies depending on the spermatozoa preparation method and whether these potential differences affect the fertilization rate of ooctyes, embryo development, embryo implantation, and pregnancy outcome. We will try to elucidate which epigenetic changes in investigated regions/genes could be used as a diagnostic tool and if they could be used as a prognostic factor for the in vitro fertilization procedure outcome. Using PCR we will analyze regions/genes which are known from the literature as specific locations where epigenetic changes occur in germ cells (in terms of maternal and paternal imprinting). As an additional part of this analysis, real-time PCR will be used to verify the impact of non-apoptotic spermatozoa selection (MACS procedure) on the expression level of genes typically involved in apoptosis.
As an additonal evaluation of effectiveness of selection method, after completion of the ICSI procedure, we will immediately check the sperm DNA fragmentation and mitochondrial membrane potential in both populations of spermatozoa (in MACS-selected and non-selected population). Studies show these factors are important indicators of sperm quality, because the analysis of DNA fragmentation shows the proportion of spermatozoa with breaks in DNA chain, and mitochondrial membrane potential shows the activity and energetic status of spermatozoa. We expect that the MACS method will improve the quality of spermatozoa population from the aspect of these two criteria, which are unfortunately not used in daily routine practise for sperm examination in all patients, although they are clinically very important factors in assessment of sperm quality.
Significance for science
The selection of a good quality spermatozoon for injection into an oocyte is a key factor for success in IVF treatment. Since the selection of spermatozoa is based mainly on its morphology, which is not always necessarily correlated with quality, it is important to use additional methods to increase the probability for selection of the highest quality spermatozoa for the IVF procedure. One such approach is non-apoptotic spermatozoa selection using the MACS procedure. Therefore, on the basis of this new procedure we expect to improve the outcome of the in vitro fertilization procedure in a selected group of infertile couples. For this approach we will determine exactly how the quality and developmental dynamic of embryos changes and what influence it has on pregnancy and miscarriage rates. This pioneering research will study how the MACS procedure affects the epigenetic status of selected spermatozoa compared to spermatozoa obtained with the standard preparation method. In addition, we will explore the correlation of the spermatozoa epigenetic status alongside quality and developmental dynamics of embryos, as well as with the pregnancy rate and miscarriages. The knowledge obtained from this study will help to explain to couples who are faced with the severest forms of infertility why IVF treatment could be less effective for them compared to other groups of infertile patients. We expect the findings about epigenetics in spermatozoa will, apart from being used as a diagnostic tool, serve also as prognostic tool or a model to predict the success of the IVF procedure in couples even before they start with infertility treatment.
Significance for the country
Due to the expected improved results of the IVF procedure, the patients’ treatment will be of better quality and more effective. This will lead to improved quality of life for patients, since it is known that infertility treatment is very stressful for such couples. Consequently, improved results will reduce the direct and indirect costs associated with the treatment of infertility, where upon improved results the couples will be less likely to need to repeat the treatment. Acquired basic knowledge of the epigenetic status of spermatozoa will be completely new for Slovenia as well as throughout the world. It will enable entrepreneurship to develop efficient products and procedures which will in turn help the patients and professionals who deal with infertility treatment with decision-making about the treatment and in the prediction of the effectiveness of such treatment.
Most important scientific results
Final report
Most important socioeconomically and culturally relevant results
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