Cíl studie: Metoda aktivace spermií je moderní metodický přístup, který se v praxi používá stále více. Neustále přibývá nových studií zaměřených na metody umělé aktivace motility lidských spermií. Standardní metody výběru spermií mohou v některých případech selhat mimo jiné i proto, že jsou izolovány spermie velice mladé, které ještě nedokončily svůj vývoj. V těchto případech může mít umělá stimulace jejich pohybu pozitivní efekt a velice usnadnit a urychlit proces výběru vhodných spermií. Jako aktivační činidla se nejčastěji využívají methylxanthiny. Názory na bezpečnost použití těchto látek na spermie však nejsou jednotné. Cílem práce je prezentovat současné poznatky o umělé aktivaci motility spermií na in vitro fertilizaci a následný embryonální vývoj. Metodika: Rešerše relevantní literatury v databázích Web of Science, Scopus, PubMed/Medline. Výsledky a závěr: Z literární analýzy vyplývá, že je tato metoda bezpečná a účinná při výběru nepohyblivých spermií. Byly provedeny vědecké studie zaměřené na ověření bezpečnosti a spolehlivosti této metody. Závěrem těchto studií je pozitivní dopad tohoto způsobu výběru především u případů spermií získávaných z varletní tkáně po metodě TESE (testicular sperm extraction). V těchto případech metoda umělé aktivace spermií usnadnila a zrychlila výběr spermií před intracytoplazmatickou injekcí spermie. Aktivovány byly spermie nepoškozené, které jsou nepohyblivé z důvodu nedokončení své maturace.
Aim: The sperm activation method is a modern methodological approach that is used more and more often in practice. The number of studies focused on methods of artificial activation of human sperm motility are constantly increasing. Standard sperm selection methods can fail in some cases, among other things, because very young sperm are isolated that have not yet completed their development. In these cases, artificial stimulation of their movement can have a positive effect and greatly facilitate and faster the process of selecting suitable sperm. Methylxanthines are most often used as activating agents. However, opinions on the safety of using these substances on sperm are not uniform. The aim of the thesis is to present current knowledge about artificial activation of sperm motility for in vitro fertilization and subsequent embryonic development. Methodology: Research of relevant literature in Web of Science, Scopus, PubMed/Medline databases. Results and conclusion: The literature analysis shows that this method is safe and effective in the selection of immotile spermatozoa. Scientific studies have been conducted to verify the safety and reliability of this method. The conclusion of these studies is the positive impact of this method of selection, especially in cases of sperm obtained from testicular tissue after method testicular sperm extraction. In these cases, the method of artificial sperm activation facilitated and accelerated the selection of sperm before intracytoplasmic sperm injection. Undamaged spermatozoa, which are immobile due to incomplete maturation, were activated.
Exposure to bisphenols has been found to have adverse effects on male reproductive function in animals. Human exposure to bisphenols is widespread. Bisphenol A (BPA) and its analogues, including bisphenol S (BPS), bisphenol F (BPF), and bisphenol AF (BPAF) are utilized in various consumer products such as food contact materials and dental resins. The effects of these compounds on male fertility and spermatogenesis are unclear and findings from human studies are inconsistent. In this cross-sectional study, we evaluated the influence of BPA, BPS, BPF, BPAF (BPs) measured in semen on number of spermatozoa, total motility, progressive motility, morphology, and DNA fragmentation. We also examined the association of bisphenols (BPs) exposure with patients' occupation. A total of 358 patients aged 17-62 years with BMI 18-42 were included in the study from 2019 to 2021. BPs were extracted using solvent extraction followed by preconcentration step and determined by high-performance liquid chromatography and tandem mass spectrometry (LC/MSMS). Bisphenols were detected in 343 from 349 analysed samples (98.3% of all the samples). In 6 samples, the concentration of all BPs was under the limit of detection and in 20 samples under the limit of quantification. We did not find a statistically significant relationship between occupation and BPs. However, we observed significant correlations between the concentration of BPA and a lower motility and normal morphology. For BPS, a significant correlation with a lower ejaculate volume and a lower total sperm count was found. BPF and BPAF were detected only in 14.3% and 23.9% of samples, respectively. For BPF and BPAF, no significant correlations with spermiogram parameters were observed. Our results show that BPs are widespread in the male population (more than 90% of analysed samples), independently of an occupation and in case of BPA and BPS having a negative impact on spermiogram parameters.
- MeSH
- benzhydrylové sloučeniny * toxicita analýza MeSH
- fenoly * MeSH
- fluorokarbony * MeSH
- lidé MeSH
- průřezové studie MeSH
- sperma * MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
BACKGROUND: Human embryonic stem cells (hESCs) have the unique ability to differentiate into any cell type in the human body and to proliferate indefinitely. Cell therapies involving hESC have shown very promising results for the treatment of certain diseases and confirmed the safety of hESC-derived cells for humans. They are used in cell therapy, mainly in targeted therapy of diseases that are currently incurable. OBJECTIVES: The aim of this study was the derivation of clinical-grade hESCs usable in drug development, non-native medicine and cell therapy. MATERIAL AND METHODS: Embryos were thawed, cultivated to the blastocyst stage if necessary, and assisted hatching was subsequently performed. Embryoblasts were mechanically isolated using narrow needles. Each line was kept as a separate batch. The derived hESCs were cultured under hypoxic culture conditions (5% O2, 5% CO2, 37°C) in a NutriStem® hPSC XF Medium with a daily medium change. RESULTS: From January 2018 to July 2020, 138 selected clients were asked for consent to donate embryos, of whom 52 did not respond, 19 terminated the storage of their embryos and 29 extended the storage. Only 38 clients (27.5%) agreed to donate embryos for the derivation of hESCs. At the same time, personal communication with clients took place and another 17 embryo donors were recruited. A total of 160 embryos from 55 donors aged 26-42 years were collected. The embryos were frozen at the blastocyst (33.1%) or morula (46.3%) stage. After the preparation of 64 embryos, embryoblasts were isolated and cultured. Finally, 7 hESC lines were obtained, 4 research-grade and 3 clinical-grade, the first in the Czech Republic. CONCLUSIONS: We established a current good manufacturing practice (cGMP)-defined xeno-free and feeder-free system for the derivation, culture and banking of clinical-grade hESC lines that are suitable for preclinical and clinical trials. The quality control testing with criteria concerning sterility, safety and characterization according to cGMP ensured the clinical-grade quality of hESC lines.
- MeSH
- buněčné linie MeSH
- embryo savčí MeSH
- embryonální kmenové buňky * MeSH
- kvalita života * MeSH
- lidé MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
Human embryonic stem cells (hESCs) have unique abilities that enable their use in cell therapy, disease modeling, and drug development. Their derivation is usually performed using a feeder layer, which is undefined and can potentially cause a contamination by xeno components, therefore there is a tendency to replace feeders with xeno-free defined substrates in recent years. Three hESC lines were successfully derived on the vitronectin with a truncated N-terminus (VTN-N) in combination with E-cadherin in xeno-free conditions for the first time, and their undifferentiated state, hESC morphology, and standard karyotypes together with their potential to differentiate into three germ layers were confirmed. These results support the conclusion that the VTN-N/E-cadherin is a suitable substrate for the xeno-free derivation of hESCs and can be used for the derivation of hESCs according to good manufacturing practices.
- MeSH
- buněčná a tkáňová terapie MeSH
- kadheriny genetika MeSH
- lidé MeSH
- lidské embryonální kmenové buňky * MeSH
- obchod MeSH
- vitronektin MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The compounds of seminal plasma have great potential as biomarkers of male fertility and can be used as a diagnostic tool for types of azoospermia. Azoospermia occurs in approximately 1% of the male population, and for an effective therapy of this form of male infertility, it is important to distinguish between obstructive and non-obstructive azoospermia. Proteins in seminal plasma can serve as biomarkers for diagnosing azoospermia. Considering the various types of obstructions, a combination of multiple proteins is advisable for diagnostic purposes. In this context, testicular and epididymal proteins are particularly significant, as they are specific to these tissues and typically absent in ejaculate during most obstructions. A combination of multiple biomarkers is more effective than the analysis of a single protein. This group of markers contains TEX101 and ECM1 proteins, combined detections of these two bring a diagnostic output with a high sensitivity and specificity. Similar results were observed for combined detection of TEX101 and SPAG1. The effective using of specific biomarkers from seminal plasma can significantly improve the existing approaches to diagnosis of the causes of male infertility.
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
(1) Objective: Breast cancer is the most common cancer in women, and the incidence of the disease continues to increase. The issue of immediate breast reconstruction (IBR) in women with BRCA mutations and breast cancer is highly topical. This study is based on the long-term experience of our workplace with the diagnosis and treatment of breast cancer in women. We use the possibilities of oncoplastic surgery, including IBR. Our effort involves learning about women's awareness of IBR with a mastectomy at the same time. (2) Methods: The method of quantitative research of women's awareness using a structured anonymous questionnaire was chosen. Out of the total number of 84 respondents who already underwent IBR, 36.9% were due to BRCA mutations, and 63.1% were due to breast cancer. (3) Results: All of the respondents learned about the possibility of IBR before treatment or during treatment planning. The information was first obtained mainly from an oncologist. Women obtained the most information regarding IBR from a plastic surgeon. Before the mastectomy, all of the respondents already knew what IBR meant, as well as about the payment of IBR by the health insurance company. All of the respondents would choose the IBR option again. A total of 94.0% of women cited preservation of body integrity as a reason for undergoing IBR, and 88.1% of women knew about the possibility of performing IBR with their own tissues. (4) Conclusions: There are few specialized centers with a team of experts in reconstructive breast surgery in the Czech Republic, especially those that perform IBR. Research has shown that all of the patients were well informed about IBR, but the vast majority only learned about IBR before the surgical procedure was planned. All of the women wished to maintain body integrity. Our study results in the recommendations for patients and for healthcare management.
- Publikační typ
- časopisecké články MeSH
Bisphenol S (BPS), the main replacement for bisphenol A (BPA), is thought to be toxic, but limited information is available on the effects of Bisphenol S on ovarian follicles. In our study, we demonstrated the presence of Bisphenol S in the follicular fluid of women at a concentration of 22.4 nM. The effect of such concentrations of Bisphenol S on oocyte maturation and subsequent embryo development is still unknown. Therefore, we focused on the effect of Bisphenol S on in vitro oocyte maturation, fertilization, and embryo development. As a model, we used porcine oocytes, which show many physiological similarities to human oocytes. Oocytes were exposed to Bisphenol S concentrations similar to those detected in female patients in the ART clinic. We found a decreased ability of oocytes to successfully complete meiotic maturation. Mature oocytes showed an increased frequency of meiotic spindle abnormalities and chromosome misalignment. Alarming associations of oocyte Bisphenol S exposure with the occurrence of aneuploidy and changes in the distribution of mitochondria and mitochondrial proteins were demonstrated for the first time. However, the number and quality of blastocysts derived from oocytes that successfully completed meiotic maturation under the influence of Bisphenol S was not affected.
- Publikační typ
- časopisecké články MeSH
Cervical mucus (CM) is a viscous fluid that is produced by the cervical glands and functions as a uterine cervix plug. Its viscosity decreases during ovulation, providing a window for non-invasive sampling. This study focuses on proteomic characterization of CM to evaluate its potential as a non-invasively acquired source of biomarkers and in understanding of molecular (patho)physiology of the female genital tract. The first objective of this work was to optimize experimental workflow for CM processing and the second was to assess differences in the proteomic composition of CM during natural ovulatory cycles obtained from intrauterine insemination (IUI) cycles and in vitro fertilization (IVF) cycles with controlled ovarian hyperstimulation. Proteomic analysis of CM samples revealed 4370 proteins involved in processes including neutrophil degranulation, cellular stress responses, and hemostasis. Differential expression analysis revealed 199 proteins enriched in IUI samples and 422 enriched in IVF. The proteins enriched in IUI were involved in phosphatidic acid synthesis, responses to external stimulus, and neutrophil degranulation, while those enriched in IVF samples were linked to neutrophil degranulation, formation of a cornified envelope and hemostasis. Subsequent analyses clarified the protein composition of the CM and how it is altered by hormonal stimulation of the uterus.
- MeSH
- biologické markery MeSH
- cervikální hlen * MeSH
- fertilizace in vitro MeSH
- lidé MeSH
- proteom MeSH
- proteomika MeSH
- umělá inseminace * MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Cíl: Endometrióza je chronické onemocnění s relativně vysokou prevalencí v ženské populaci. Jak nemoc samotná, tak i její chirurgická léčba mohou nepříznivě ovlivnit plodnost pacientek. Z toho důvodu se endometrióza nabízí jako možná indikace k ochraně reprodukce metodami kryoprezervace. Cílem práce je prezentovat současné poznatky o možnostech ochrany reprodukce v této subpopulaci. Metodika: Rešerše relevantní literatury v databázích PubMed/Medline, Web of Science a Scopus. Výsledky: Ochrana reprodukce metodami kryoprezervace se dosud využívala zejména v péči o onkologicky nemocné ženy. S narůstajícími zkušenostmi se výrazně zvýšila efektivnost i dostupnost těchto metod a indikační spektrum bylo rozšířeno na vybraná benigní onemocnění, jako je právě endometrióza. V současnosti jsou v praxi zavedené tři techniky: kryoprezervace embryí, kryoprezervace oocytů a kryoprezervace ovariální tkáně. Nejčastěji se využívá kryoprezervace oocytů, která je pro pacientku nejvýhodnější a dle dostupných dat představuje efektivní způsob, jak rizikovým pacientkám s endometriózou zvýšit šanci na budoucí graviditu. Smyslem je realizace ochrany reprodukce před plánovaným operačním výkonem. Závěr: Dia gnóza endometriózy negativně ovlivňuje fertilitu žen. Pro část pacientek je řešením ochrana reprodukce metodami kryoprezervace. K definování přesných, prakticky aplikovatelných indikačních kritérií, potenciálních rizik postupů a jejich benefitů i nákladové efektivnosti je potřeba dalších klinických studií.
: Objective: Endometriosis is a chronic disease with a relatively high prevalence in the female population. Both the disease itself and its surgical treatment can adversely affect the fertility of patients. For this reason, endometriosis is offered as a possible indication for fertility preservation by cryopreservation methods. The aim of this paper is to present the current knowledge on the options of fertility preservation in this subpopulation. Methods: Search of relevant literature in PubMed/Medline, Web of Science and Scopus databases. Results: Fertility preservation by cryopreservation methods has so far been used mainly in the care of women with cancer. With increasing experience, the effectiveness and availability of these methods have increased significantly and the indication spectrum has been extended to selected benign diseases such as endometriosis. Three techniques are currently established in practice: embryo cryopreservation, oocyte cryopreservation and ovarian tissue cryopreservation. Oocyte cryopreservation is the most commonly used technique, since it is the most advantageous for the patient and, according to the available data, is an effective way to increase the chances of future pregnancy for patients with endometriosis The purpose is to realize the protection of reproduction before the planned operation. Conclusion: The diagnosis of endometriosis negatively affects the fertility of women. For some patients, the solution is fertility preservation by cryopreservation methods. Further clinical studies are needed to define exact, practically applicable indication criteria, potential risks of procedures and their benefits and cost-effectiveness.
Cíl: V současné době rychle přibývá nových studií o asistované aktivaci oocytů, která může výrazně zefektivnit celý proces in vitro fertilizace. Oplození oocytů konvenční metodou i pomocí intracytoplazmatické injekce spermie může selhat z důvodu nedostatečné aktivace oocytu. Důvodem bývají zejména odchylky v enzymovém vybavení spermií nebo oocytů či nefunkční aktivační kaskáda. V řadě případů lze oplození dosáhnout užitím umělé aktivace oocytů pomocí aplikace donorů vápníkových iontů na oocyty po mikroinjekci spermie. Názory na bezpečnost a spolehlivost této metody však nejsou jednotné. Cílem práce je prezentovat současné poznatky o asistované aktivaci oocytů a o jejím dopadu nejen na in vitro fertilizaci, ale také na následný embryonální a fetální vývoj. Metodika: Rešerše relevantní literatury v databázích Web of Science, Scopus a PubMed/Medline. Výsledky a závěr: Z literárních údajů i vlastních zkušeností autorů vyplývá, že je tato metoda účinná a z hlediska dalšího vývoje embrya, plodu i postnatálního vývoje bezpečná. Byly provedeny rozsáhlé metaanalýzy zaměřené na tuto metodu, které nezjistily negativní dopad nejen na embryonální a fetální vývoj jedince, ale tato metoda neměla negativní dopad ani na psychosomatický vývoj narozených dětí.
Objective: Currently, there is a rapid increase in studies on assisted oocyte activation, which can significantly improve the process of in vitro fertilization. Fertilization of oocytes by conventional methods and by intracytoplasmic sperm injection can be affected by insufficient activation of the oocyte. The reason is mainly deviations in the enzymatic equipment of sperm or oocytes or a non-functional activation cascade. In many cases, fertilization can be achieved using artificial oocyte activation by applying calcium ion donors to the oocytes after sperm microinjection. However, opinions on the safety and reliability of this method are not uniform. The aim of the thesis is to present current knowledge about assisted oocyte activation and its impact not only on in vitro fertilization, but also on subsequent embryonic and fetal development. Methodology: Research of relevant literature in Web of Science, PubMed/Medline and Scopus databases. Results and conclusions: Based on the literature data and the authors’ own experience, it follows that this method is effective and safe from the point of view of further development of the embryo, fetus and postnatal development. Extensive meta-analyses focused on this method were carried out, which did not find a negative impact not only on the embryonic and fetal development of the individual, but this method did not have associated with a negative impact on the psychosomatic development of the children.
- MeSH
- fertilizace in vitro * metody MeSH
- intracytoplazmatické injekce spermie MeSH
- ionofory kalciové aplikace a dávkování MeSH
- lidé MeSH
- oocyty * fyziologie MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- přehledy MeSH
