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Інституційний репозитарій ІПКіК НАН України — відкрита електронна платформа для збереження та поширення наукових досліджень у галузі кріобіології та кріомедицини..

Документів у репозитарії: 9

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Metabolic component of adaptation to cold stress in in vivo and in vitro experiments
Myrnyi Vladyslav
;
Moisieieva Nataliia
(Bulletin of problems in biology and medicine –, 2025)
In the present work, a comprehensive study of metabolic and cellular responses to cold stress in in vivo and in vitro models was conducted, allowing the assessment of the effect of the synthetic neuropeptide dalargin on the formation of adaptive metabolic and cytoprotective responses. It was found that administration of dalargin significantly increased blood triglyceride concentrations, which were reduced under chronic cold stress, while the total cholesterol concentration remained decreased. This dynamic indicates the selective action of the peptide on rapidly mobilized energy lipids, whereas cholesterol is regulated by complex, long-term synthesis and degradation mechanisms. It is emphasized that maintaining triglyceride levels is critically important for ensuring the energy homeostasis of the organism under cold stress conditions. In vitro, cold stress was found to reduce the mitotic index and viability of fibroblasts, whereas dalargin partially prevents the development of these negative changes. Integration of in vivo and in vitro data provides grounds to conclude that dalargin acts as a selective metabolic and cytoprotective agent, ensuring rapid adaptation of lipid and energy metabolism, maintaining cellular activity and viability under stress conditions, and contributing to the stabilization of systemic homeostasis. The results highlight the potential of dalargin as an effective modulating factor for supporting metabolic and cellular adaptation under cold stress.
Effect of chronic cold stress and dalargin on blood parameters in gunea pigs
Myrnyi Vladyslav
;
Moisieieva Nataliia
(Problems of Cryobiology and Cryomedicine, 2025)
Th is study investigates the eff ects of the synthetic neuropeptide Dalargin on hematological parameters and blood electrolyte homeostasis in guinea pigs under conditions of chronic cold stress (CCS) at 4 °C over a fi ve-day period. It was found that CCS led to a reduction in platelet count and disruption of the leukocyte profi le, including an increase in band neutrophils accompanied by a decrease in segmented neutrophils, lymphocytes, and monocytes, as well as a decline in the adaptation index. Simultaneously, elevated concentrations of calcium, potassium, and chloride were observed, indicating disturbances in electrolyte balance and ionic homeostasis. Dalargin administration prevented these changes, promoting the restoration of physiological leukocyte ratios, increasing platelet counts, and normalizing key plasma electrolyte concentrations. Moreover, an increase in the adaptation index was recorded, indicating activation of the hematopoietic system and mobilization of the organism’s adaptive potential. Th ese fi ndings support the potential of dalargin as an eff ective agent for maintaining blood system integrity, electrolyte balance, and adaptive capacity under CCS conditions.
Preservation of protein fraction and hormones of placental blood serum after freeze - drying
Shevchenko Mariia
;
Skoryk Anastasiia
;
Salnykov Dmytro
;
Gorina Olga
;
Ivanov Yevhen
(Problems of Cryobiology and Cryomedicine, 2026)
Modern medicine, especially in military and emergency settings, requires eff ective and logistically convenient dosage forms suitable for use at all stages of hospitalization. Lyophilized preparations meet these requirements, as they are characterized by high physical and chemical stability, a long shelf life, no need for special storage conditions; their minimal weight and volume facilitate transportation, while their rapid rehydration ensures ease of use [32]. Traumatic brain injury (TBI) complicated by hypothermia is a common problem among military personnel and civilians in combat zones, particularly in Ukraine. Th is is due to the fact that injuries oft en occur in conditions of low temperatures, the absence of adequate shelter, and complicated or delayed evacuation [6, 30]. A signifi cant proportion of victims sustain injuries against the background of general hypothermia, as evidenced by the consistent incidence of hypothermia among the wounded [6, 10, 25, 30]. Hypothermia can signifi - cantly exacerbate the consequences of TBI, contributing to the development of infl ammatory processes, coagulopathy, microcirculatory disorders, and an increased risk of secondary brain injury, making this combination one of the most dangerous clinical conditions in modern warfare. Furthermore, hypothermia is an independent risk factor for mortality following combat-related injuries, being associated with increased mortality [6, 30] and poorer clinical outcomes in trauma patients [25, 29, 31]. Given the above, the current task is to improve existing treatment approaches and search for innovative multitargeted therapy methods. One of the modern biomedical approaches to the eff ective restoration of damaged organs and regulatory systems is the use of placental derivatives, particularly placental blood components such as cells, plasma, serum, etc. Th ese components possess unique therapeutic potential and play a critical role in the regeneration of damaged tissue by suppressing infl ammatory reactions and apoptosis, inducing angiogenesis, and activating regenerative processes [19, 22, 23, 27]. Placental derivatives have been shown to possess immunomodulatory properties [16] and demonstrate effi cacy in the treatment of heart failure [21]. Th ey have also been found to improve the course of experimental encephalomyelitis [26], ischemic stroke, experimental Parkinson’s disease, and spinal cord injuries [13—15], stimulate the regeneration of nervous tissue and exhibit neuroprotectiveproperties [11]. Th ese studies underscore the potent multitargeted therapeutic eff ect of placental derivatives, which is mediated through reparative and regulatory processes. The unique profile of biologically active substances of placental blood serum (PBS), including proteins, hormones, cytokines, growth factors, neurotrophic factors and immunomodulators, determines its regenerative, anti-infl ammatory, immunosuppressive, anti-apoptotic and neuroprotective properties [17, 23]. Th is provides a rationale for the potential effi cacy of PBS in mitigating the consequences of concussive injury complicated by hypothermia. Given the signifi cant therapeutic potential of PBS, ensuring the long-term preservation of its biological properties remains a primary objective. Freeze-drying, as one of the most eff ective methods for preserving PBS, can potentially ensure the maximum stability of its components. However, the effi cacy of this process largely depends on the initial composition of the source material. It is well established that PBS composition is characterized by high variability and is infl uenced by several factors, particularly the clinical features of pregnancy and labor [5, 8, 20, 24]. This necessitates a rigorousselection of placental blood samples for further experimental and clinical use, taking into account the maternal medical history, the course of pregnancy and childbirth, and the presence of gestationalcomplications. Thus, given that both the course of pregnancy and freeze-drying can aff ect the content of biologically active compounds in the PBS, which are potentially capable of having a corrective eff ect in TBI against the background of hypothermia, the aim of the work was to study the safety of placental blood serum aft er freeze-drying, taking into account the course of pregnancy in parturient women.
Comprehensive characterization of human Wharton’s jelly mesenchymal stromal cells isolated by the combined explant-enzymatic method
Obushko Ruslana
;
Trufanova Natalia
;
Revenko Olena
;
Cherkashina Daria
;
Lipko O.
(2026)
Human mesenchymal stromal cells (MSCs) derived from Wharton’s jelly (WJ) are a premier source for regenerative medicine. However, standardized isolation protocols that ensure both high cell yield and maintained proliferative potential remain a subject of active research. This study is aimed to evaluate the efficiency of a combined explant-enzymatic method for human WJ-MSC isolation and characterize the resulting cell populations. Materials and methods. Cells were isolated using a combined explant-enzymatic method. Minced WJ fragments were cultured on gelatin-coated surfaces for 7 days, then explants underwent a brief (5–10 min) in situ enzymatic treatment with a trypsin-Versene solution, followed by further co-cultivation of the tissue fragments and released cells for an additional 7 days prior to the first passage. Cell characterization included colony-forming unit (CFU), immunophenotyping (CD29, CD90, CD73, CD105, CD34, CD45), directed differentiation (adipogenic and osteogenic), and secretory activity assays. Proliferative kinetics were assessed by calculating the population doubling time (PDT) across 5 passages. Secretory profile of WJ-MSCs (growth factors and cytokines regulating reparative-regenerative processes) was studied by enzyme-linked immunosorbent assay of conditioned media collected after 48 h cell incubation in minimal culture media. Results. The combined method ensured a stable cell yield with high reproducibility. Significant inter-donor variability in primary cell yield was noted. Isolated cells fulfilled the ISCT minimal criteria, exhibiting typical fibroblast-like morphology, characteristic immunophenotype, differentiation potential, and a high proportion of CFU, indicating the preservation of primitive progenitors. Proliferative analysis showed that PDT values remained comparable and did not change significantly between passages 1 and 3. Conditioned media from WJ-MSCs contained wide range of growth factors and cytokines, with inter-donor variability: VEGF, EGF, PDGF-BB, SCF, TNF-α, and TGF-β exhibited wide fluctuations, whereas NGF-β and FGF-β were relatively stable. Overall, WJ-MSC CM were characterized by very high levels of VEGF and EGF, high PDGF-BB, elevated TNF-α and TGF-β, and the lowest levels of FGF-β. Conclusions. The combined explant-enzymatic method is an efficient and standardized protocol for human WJ-MSC.
Ontogenetic stage affects rat neural cell spheroids formation and properties
Sukach Oleksandr
;
Maiorova Olga
;
Ochenashko Olga
;
Vsevolodska Svitlana
(2026)
Aim. To compare the formation efficiency and morphometric, mechanical, and morphofunctional properties of neural spheroids derived from brain cells of rat embryos (embryonic day 15, E15) and neonatal rats (postnatal day 0, P0). Methods. Neural cells were isolated by mechanical-enzymatic dissociation, and spheroids were formed by the hanging drop method. Spheroid formation efficiency, diameter, sphericity, mechanical stability, and cell behavior after spheroid transfer to an adhesive substrate were evaluated. Results. Embryonic brain-derived cells demonstrated higher spheroid formation efficiency, producing larger, more spherical structures with less size variability and greater mechanical sta- bility than neonatal cells. After attachment, E15 spheroids rapidly disintegrated, accompanied by intense radial cell migration and the formation of a confluent layer exhibiting signs of neuronal and glial differentiation. In contrast, neonatal P0 spheroids maintained a compact 3D structure and formed stable neuronal networks. Conclusions. The ontogenetic stage of neural cells significantly affects spheroid self-organisation, mechanical stability, and morphofunctional properties, highlighting its importance in selecting cell sources for 3D neural tissue models.