Meiotic errors during oocyte maturation are the primary cause of embryonic aneuploidy. Coordinated functioning of the cytoskeleton and its associated contractile proteins is essential for karyokinesis and cytokinesis. This review summarizes studies investigating the roles of actin, myosin, and kinesin in mouse and human oocyte maturation. Mechanisms underlying the involvement of contractile proteins in oocyte maturation are described. Specifically, actin participates in spindle assembly during the first and second meiotic divisions by stabilizing K-fibers. Cytoplasmic actin, together with myosin-5b and myosin-2, mediates the migration and orientation of the meiotic spindle to the periphery, driving asymmetry in human oocytes; in mouse oocytes, actin and myosin-5b are also responsible for central positioning of the nucleus in prophase I. Polar body extrusion occurs via contraction of the actomyosin contractile ring. Kinesins are also widely represented in oocyte maturation processes. In mouse oocytes, Kif11, Kif16A, and Kif16B contribute to spindle formation and stabilization, whereas KifC1 stabilizes kinetochore fibers (K-fibers). Additionally, Kif16A and Kif16B regulate cell cycle progression by modulating spindle assembly checkpoint activation and the G2–M meiotic transition, respectively. In human oocytes, HSET (kinesin-14) is responsible for spindle pole determination, and Kid (kinesin-10) is involved in chromosome alignment at the metaphase plate.
Objective: The aim of this study is to compare the prognostic significance of the KIDScore, PGT-A, and MitoScore assessment systems in relation to the probability of embryo implantation at the blastocyst stage, pregnancy outcomes, and obstetric complications in ART cycles. Materials and Methods: The study conducted a retrospective analysis of embryological protocols and the results of PGT-A. In euploid embryos, the mitochondrial DNA MitoScore was assessed. The clinical outcomes of the resulting pregnancies were evaluated. Results: It was found that higher KIDScore values were more often determined in euploid embryos. When analyzing the relationship between KIDScore and MitoScore, no statistically significant dependence was found. However, there was a tendency towards a higher frequency of low and medium MitoScore values among embryos in groups with medium and high KIDScore values. In our study, we found a statistically unconfirmed tendency towards a higher frequency of clinical pregnancies and live births in the group of patients who underwent embryo transfer with high KIDScore and medium MitoScore values. There was an increase in the percentage of complicated pregnancies after embryo transfer with low KIDScore values. In our study, the most common obstetric complications were preeclampsia and gestational diabetes mellitus. Conclusions: The obtained data allow us to consider the cultivation of embryos using video surveillance in combination with the assessment of the implantation potential of embryos based on artificial intelligence technologies as a non-invasive tool for predicting the chromosomal status of the embryo, as well as to use this technique as an additional tool when selecting an embryo for transfer into the uterine cavity. Proper integration of the KIDScore, PGT-A, and MitoScore embryo assessment systems can help improve the accuracy of selecting an embryo with the highest potential for implantation, which is crucial for implementing a personalized approach to patients and transferring a single embryo to prevent multiple pregnancies and their associated complications.
Background: Elevated levels of sperm DNA fragmentation can negatively affect outcomes of assisted reproductive technology programs, making the selection of spermatozoa with intact genetic material a critical step. Microfluidic technologies can provide selection of spermatozoa with low levels of DNA damage. Objective: To compare the DNA fragmentation index in spermatozoa from native ejaculate, after density gradient centrifugation, and after microfluidic selection (SPRINT chip) using a Sperm-Cyto flow cytometer. Materials and methods: The study included 25 men of reproductive age (mean age 37 years). Ejaculate samples were analyzed according to standard protocols and divided into groups: native sample, spermatozoa after density gradient centrifugation, and spermatozoa after selection on the SPRINT microfluidic chip. In each group, the DNA fragmentation index was assessed using the SCSA (Sperm Chromatin Structure Assay) method with a Sperm-Cyto flow cytometer. Results: Microfluidic selection provided the selection of spermatozoa with a low DNA fragmentation index (2.30 ± 1.78
Azoospermia is defined as the absence of spermatozoa in the ejaculate after centrifugation. This condition represents one of the most severe causes of male infertility. The prevalence of azoospermia is up to 1
Background: Infertility represents a global medical and social problem, and endometrial dysfunction (thinning, impaired receptivity, synechiae, etc.) remains one of the key causes of implantation failure and limitation of the effectiveness of assisted reproductive technology (ART) programs. In this regard, new pathogenetic approaches are being actively investigated, among which cell technologies occupy a special place. Objective: To analyze current scientific literature data on the potential of mesenchymal stromal cells derived from menstrual blood (MenSCs) for restoring functional activity of the endometrium and improving the effectiveness of ART programs. Materials and methods: This review summarizes the results of fundamental research, preclinical trials on animal models, and early clinical studies on the isolation, cultivation, phenotyping, and transplantation of human MenSCs. The review includes data from foreign and Russian articles found in Pubmed, Elsevier, and Cochrane Library databases on this topic, published over the last 7 years. Results: Literature analysis confirms that menstrual blood is an accessible and reproducible non-invasive source of MSC cells with a phenotype and properties consistent with the criteria of the International Society for Cell Therapy (ISCT). MenSCs demonstrate high proliferative potential, multilineage differentiation capacity, and pronounced paracrine activity directed toward angiogenesis and modulation of endometrial receptivity. Based on recent studies, the safety profile of MenSCs is assessed as favorable. Conclusion: Further research in this area is needed to standardize protocols for cell isolation and transplantation, and ultimately to use optimized methods in clinical practice of ART programs to increase implantation and pregnancy rates.
Folliculogenesis is a complex process culminating in the formation of the oocyte-cumulus complex (OCC). This review examines the stages of OCC growth and development from the primordial follicle to the preovulatory stage and describes the characteristics of nuclear and cytoplasmic oocyte maturation, with an emphasis on its reliance on cumulus and mural granulosa cell differentiation. In clinical embryology, follicular fluid obtained during transvaginal puncture—which contains cumulus and granulosa cells—is routinely discarded; however, it harbors biomarkers that enable the assessment of oocyte and future embryo potential. Understanding the mechanisms of communication between the oocyte and its somatic environment provides fundamental insights, enables the refinement of cellular technologies, aids in developing personalized approaches to infertility treatment, and ultimately enhances the efficacy of assisted reproductive technologies (ART). Special attention is given to the transzonal projections (TZPs) of cumulus cells, bidirectional communication via gap junctions, and the role of amphiregulin (AREG) in activating the MAP kinase cascade during meiotic arrest resumption. We demonstrate how fundamental knowledge of the oocyte microenvironment can be harnessed to design cellular technologies for clinical applications.
Objective: Prenatal whole-exome sequencing (prWES) is a promising molecular diagnostic tool that can be used to clarify the genetic etiology of prenatally detected abnormalities, including cases in which standard genetic testing fails to establish their cause. However, data on its diagnostic yield and the spectrum of detectable abnormalities in Russian clinical practice remain limited. Objective: To perform a retrospective analysis of the indications for prWES, its diagnostic yield, and the spectrum of molecular findings in a Russian clinical cohort. Materials and methods: This retrospective cohort study included 485 pregnancies in which prWES was performed. Results: The most common indications for testing were fetal structural anomalies (43.71
Background: Advances in assisted reproductive technologies (ART) make it possible today not only to overcome many types of infertility, but also to preserve gametes or embryos prior to gonadotoxic treatment. In a number of cases, when urgent fertility preservation programs are required in women, immature oocyte-cumulus complexes (OCC) are retrieved with subsequent in vitro maturation (IVM). This method has become widespread across various clinical indications; however, its efficiency remains relatively low, requiring optimization of protocols and the study of factors influencing maturation. Objective: The aim of this study was to evaluate the relationship between the diameter of retrieved oocytes, the density of their cumulus cells, and the maturation rate in patients of different age groups. Methods: The study included two groups of patients: the OPU-IVM group, in which oocytes were obtained via transvaginal follicular aspiration, and the OTO-IVM group, where oocytes were harvested ex vivo from excised ovarian tissue. For all OCC, visual assessment of cumulus volume, structure, and density was performed, alongside measurement of the mean oocyte diameter. Results: The overall maturation efficiency to the MII stage was 50.6
This review analyzes several “hot topics” common to cell biology and reproductive medicine. The prospects and challenges of implementing advanced cell technologies into clinical practice in reproductive medicine are discussed. In particular, the significance of 3D cell and tissue culturing, multi-omics technologies, and stem cell-based technologies for improving the effectiveness of infertility treatment is briefly highlighted.
Background: The identification of non-invasive markers for selecting the most promising embryo is extremely important today. Morphological assessment is the basic criterion for embryo evaluation in in vitro fertilization (IVF) cycles. However, data on the correlation between embryo morphology, euploidy, and implantation potential are conflicting. With the advent of time-lapse incubators, it has become possible to assess morphokinetic parameters of embryo development and to use them for predicting both ploidy and implantation potential. Based on the morphokinetics of day 5–6 embryos, the computer software of time-lapse incubators provides an automated assessment of embryo implantation potential (KIDScoreD5). This algorithm was developed taking into account the currently known morphokinetic parameters most significant for embryo implantation: cleavage uniformity, blastocyst formation time, and the quality of the inner cell mass and trophectoderm. KIDScoreD5 allows ranking of embryos, which is particularly relevant when several embryos with similar morphology are available. Objective: To evaluate the relationship between KIDScoreD5 and embryo ploidy, and to assess the effectiveness of KIDScoreD5 in predicting embryo ploidy in patients of different age groups. Materials and methods: A retrospective analysis was performed on 1124 embryos with known ploidy obtained from 418 patients undergoing infertility treatment at the Skyfert Clinic in St. Petersburg from January 1, 2021, to March 31, 2025. The mean KIDScoreD5 score of day 5 and day 6 embryos that underwent PGT-A was assessed in two age groups: under 37 years and 37 years and older. The Mann-Whitney test was used for group comparisons. Spearman’s rank correlation coefficient was calculated to assess the relationship between quantitative variables. ROC analysis was used to evaluate the predictive quality of individual parameters and of our developed model for calculating the probability of embryo euploidy. Binary logistic regression was used to assess the relationship between maternal age, KIDScoreD5 score, embryo developmental day, and embryo ploidy. Results: The frequency of euploid embryos decreased significantly with increasing maternal age starting from 37 years (p < 0.0001), based on which patients were divided into two age groups: under 37 years and 37 years and older. The frequency of euploid embryos differed significantly between the two age groups (60.1 and 30.4
This brief commentary addresses aspects of the handling and use of cell products under individual treatment protocols. It highlights two potentially controversial issues: (1) the potential use of the individual pathway to administer allogeneic products without regulatory approval, and (2) the lack of well-defined regulatory criteria in this field.
Canine mammary tumors (CMTs) are the most common neoplasms in intact female dogs and represent a spontaneous, immune competent model for human breast cancer (HBC), particularly triple negative/basal like subtypes. Epidermal growth factor receptor (EGFR/ErbB1/HER1) is overexpressed in approximately 42–55
Objective: The aim of this study was to investigate the effect of heparin at a concentration of 1 IU/mL on the osteogenic differentiation potential of human adiposederived mesenchymal stem/stromal cells (ASCs) during in vitro co-cultivation. Materials and methods: The phenotypic profile of ASCs cultured with or without heparin for 14 days was assessed by flow cytometry using appropriate antibodies according to the manufacturer’s protocol on a MACSQuant cytometer. The migration and proliferation potential of ASCs in the presence of heparin was evaluated using a realtime cell monitoring system (xCELLigence ® RTCA DP). After 14 days of co-cultivation with heparin, the expression of osteogenic marker genes was assessed by real-time PCR. In addition, the differentiation profile of ASCs cultured with heparin was evaluated by alizarin red staining to detect mineralization nodules after 21 days of cultivation. Finally, the levels of growth factors, chemokines, and pro and antiinflammatory molecules were measured in the supernatants of 14-day cultures. Results: Compared to the control group, the MSC + heparin model showed a significant decrease in the percentage of cells expressing the stem cell markers CD73, CD90 and CD105; increased proliferative activity but decreased migratory activity of ASCs; increased relative mRNA expression of osteogenic genes (ALPL, RUNX2, BMP2, BMP6) and the adhesion gene CD49d; and a larger mineralization area after 21 days of cultivation. A trend towards increased secretion of VEGF and the proinflammatory cytokine IL6 was observed in the MSC + heparin model. Conclusion: These results may serve as a basis for developing new clinical strategies for the treatment of surgical patients undergoing osteosynthesis who are at high risk of thrombosis.
Objective: To develop a comprehensive approach for identifying live cell nuclei in images without fluorescent labels. Since cell biology involves counting cells, assessing cell growth dynamics, and confluence, it is expedient to automate the collection of these data. Machine learning algorithms are used for automation and must be trained on images of specific cell cultures. Training algorithms is a labor-intensive process and requires lengthy manual annotation. Additionally, available machine learning-based analysis methods have low accuracy in identifying living cells without fluorescent staining. Materials and methods: The methodology involved the use of convolutional neural networks based on an algorithm for segmenting cell nuclei in fluorescent and histological images using StarDist. To create annotated phase-contrast images of cell cultures, samples were stained with the nuclear fluorescent dye DAPI, followed by the rejection of poor-quality images using classification in the CellProfiler Analyst program. The StarDist-based model was trained on 1 130 images of automatically annotated nuclei in phase-contrast images of human respiratory tract epithelial cell cultures, obtained with a 10× lens, 1 600 × 1 200 pixels in size, and 16-bit grayscale depth. Results: The resulting model showed good accuracy (F1 = 0.765) in segmenting nuclei on the validation dataset. The model was used to determine the population doubling time of the epithelial cell culture. Conclusion: The developed approach made it possible to create annotations and train a machine learning model to obtain data without the use of fluorescent labels (“label-free”) on live cell cultures.
Intensive development of personalized medicine is opening up new possibilities for the development of regenerative medicine technologies and the translation of these developments into the clinic. One of the rapidly developing directions in creating new therapeutic approaches is the use of bioprinting for the fabrication of tissue and organ constructs. Particular attention is drawn to the development of skin equivalents capable of reproducing the complex architectural organization and functional properties of skin tissues. The review analyzes publications presented in the Scopus, PubMed, and RSCI databases, covering the fields of bioprinting, tissue engineering, and regenerative medicine. Published data were used that are devoted to the development of biomaterials, 3D-bioprinting protocols, characteristics of bioprinted skin constructs, and results of both preclinical and clinical studies, relevant as of September 2025. The analysis showed that one of the most promising directions is the optimization of 3D-bioprinting of skin constructs based on the use of fibroblasts, keratinocytes, and innovative biomaterials such as hydrogels, collagen matrices, and GelMA. These technologies enable the creation of full-thickness, vascularized structures, ensuring sufficiently high accuracy of the spatial distribution of cells and support for the microenvironment necessary for tissue regeneration. Further studies on optimization of printing parameters, proper selection of bioink components, and integration of fibroblasts and other cellular components will allow more precise modeling of the dermal layers and stimulation of regeneration processes. The application of additional biological factors will contribute to the formation of a stable vascular network and better engraftment of constructs, which will significantly enhance the functional integration of printed constructs into the recipient tissue. Thus, the integration of advanced 3D-bioprinting methods, optimized bioinks, and multicellular constructs opens prospects for creating a new generation of skin equivalents, which will not only accelerate the regeneration process but also provide an aesthetically optimal outcome for patients suffering from severe burns, injuries, and other skin damage.
Objective: This study investigated the cytotoxic and pro-apoptotic effects of hypericin on human gingival fibroblasts (HGF) under non-photoactivated conditions. Methods: HGF cells (ATCC® PCS-201-012TM) were exposed to increasing concentrations of hypericin (1–100 µM), and cell viability was assessed using the MTT assay at 24, 48, and 72 h. Long-term proliferative capacity was evaluated by clonogenic assays, while DNA synthesis was determined using BrdU/EdU incorporation. Intracellular reactive oxygen species (ROS) production was measured using the DCFH-DA fluorescent probe. Apoptotic responses were further analyzed by caspase-3/7 activity assays and Annexin V-FITC/PI flow cytometry. Results: Hypericin induced a significant, concentration- and time-dependent reduction in HGF cell viability, colony formation, and DNA synthesis. Exposure to higher concentrations (≥10 µM) resulted in a marked increase in intracellular ROS levels. In parallel, caspase-3/7 activation and apoptotic cell populations were significantly elevated in treated cells compared with controls. Importantly, these effects were observed in the absence of photodynamic activation, suggesting that hypericin may exert intrinsic cytotoxic and oxidative effects independent of light-induced ROS generation. Conclusion: Overall, hypericin demonstrates dose-dependent pro-oxidant and pro-apoptotic activity in gingival fibroblasts. These findings provide new insight into the biological responses of normal oral fibroblasts to hypericin and highlight the importance of defining safe exposure levels for potential oral and dental applications.
Objective: This study examines the influence of the lipid-soluble lignan sesamin on the physical properties of model lipid membranes. Its ability to alter the transmembrane distribution of electrical potential and the lipid packing in membranes containing neutral phosphatidylcholines (PC) and negatively charged phosphatidylserines (PS) was assessed. Methods: Electrophysiological method for determining changes in electrical potential at the membrane/aqueous solution interface and differential scanning microcalorimetry for measuring the thermodynamic characteristics of lipid vesicles. Results: Sesamin was found to reduce the membrane boundary potential regardless of membrane composition, with the effect not exceeding 20 mV. It was also shown to significantly affect the main temperature, peak width, and phase transition enthalpy of PC membranes, with the effect decreasing as the acyl chain length of the lipid increased. Furthermore, sesamin induced a decrease in the phase transition temperature of PS and the formation of additional mixed lipid–sesamin phases. Conclusion: These results suggest that sesamin reduces lipid packing density in membranes and warrant further investigation of how these effects contribute to its pharmacological activity.
Although melanoma is one of the most aggressive skin cancers with limited treatment options at advanced stages, immune checkpoint inhibitors and targeted therapies have led to meaningful clinical progress. Cancer vaccines have long been investigated as a strategy to activate anti-tumor immunity in melanoma, with early approaches primarily focused on shared tumor-associated antigens (TAAs). However, these initial efforts were constrained by limited immunogenicity, central and peripheral tolerance to self-antigens, suboptimal antigen selection, and logistical challenges. Recent advances in cancer genomics, bioinformatics, vaccine engineering, and immunotherapy have catalyzed a paradigm shift toward vaccines targeting tumor-specific antigens, particularly patient-specific neoantigens arising from somatic mutations. In this review, we summarize both historical and contemporary developments across key melanoma vaccine platforms, including peptide-based, dendritic cell, whole-cell, viral, bacterial, mRNA, and nanoparticle-based vaccines. We highlight preclinical and clinical findings that inform antigen selection, the role of adjuvants and delivery systems, and the rationale for combinatorial strategies incorporating immune checkpoint inhibitors or tumor microenvironment modulators. While whole-cell and vector-based vaccines provide broad antigenic coverage, neoantigen-based and nanoparticle-enabled vaccines offer highly specific, personalized, and potentially scalable approaches that align with precision immuno-oncology. Ongoing challenges include manufacturing complexity and cost, tumor immune evasion and resistance mechanisms, and the need for rationally designed combination regimens to maximize durable clinical benefit.
Objective: Exposure to environmental toxins can negatively impact brain health, and the underlying mechanisms are thought to be oxidative stress and neuroinflammation. Metformin is a medication used as a first-line treatment for type 2 diabetes and also has neuroprotective properties. Senescent astrocytes have been shown to accumulate with age and in the context of many neurodegenerative diseases. The present study investigated the effects of metformin on the oxidative stress-induced premature senescence in astrocyte cells. Material and methods: Primary culture astrocyte cells were pretreated with metformin before the treatment of H2O2 every 72 h. Senescence-associated beta-galactosidase (SA-β-gal) staining was performed to confirm senescence induction, and followed by mRNA expression analysis of cell cycle inhibitors (p53, p21WAF1 and p16 INK4a) and senescence-associated secretory phenotype (SASP) proteins by q-PCR. Intracellular reactive oxygen species (ROS) level was measured by DCFH-DA method. Results: H2O2 significantly increased the number of SA-β-gal positive cells and mRNA levels of cell cycle repressors (p53 and p21WAF1) and SASP proteins (IL-6, IL-1β, CXCL1, and CCL2). Metformin pretreatment significantly reduced H2O2-induced senescent cell number. H2O2–induced increase in levels of p53, p21WAF1, IL-6, CXCL1, and CCL2 mRNA and ROS was significantly inhibited by metformin. Conclusion: In the present study, we show for the first time that metformin significantly prevents premature senescence caused by oxidative stress in astrocyte cells. Metformin prevented H2O2-induced senescence development by decreasing inflammation and oxidative stress and it would be beneficial to support the mechanism of the protective role of metformin on astrocyte senescence in neurodegenerative diseases.
