Problems of Cryobiology and Cryomedicine

Hypothermic Storage of Mesenchymal Stromal Cell-based Spheroids at a Temperature of 22°C

Natalia Trufanova — 2024-12-02

Spheroids formed from mesenchymal stromal/stem cells (MSCs) provide optimal microenvironment for cell potential and stemmness support being also promising structures for relevant research models as well as cell therapies development. Practical application requires the development of effective methods for storing spheroids. Storage at ambient temperature (22°C) can be safer and more convenient for short term transportation of MSC-based spheroids. Herein we report that the spheroids formed by human adipose tissue-derived MSCs can be stored at ambient temperature (22°C) for up to 7 days with efficient preservation of viability, metabolic activity, ability to attach, and differentiation capacity. Cytoskeleton reorganization, cell size reduction, and metabolic activity decrease were shown to result from spheroidal formation and were extremely important for the development of spheroid resistance to the subsequent ambient storage.

Probl Cryobiol Cryomed 2024; 34(2):186–200

Blood Leukocyte Indices in Young and Aged Rats Under Desynchronosis Initiation Against the Background of Preventive Whole-body Cryostimulation

Viktoria Lomako — 2023-09-30

It was speculated that the whole-body cryostimulation (WBC) would adjust the negative effect of desynchronosis on the body. One-time WBC was performed in a cryochamber at –120°C a day before the desynchronosis simulation, which was initiated by shifting the light regimen. The WBC effi cacy was determined by leukocyte parameters of the blood of 6- and 18-monthold rats. Types of leukocytes were counted in blood smears. Integral leukocyte indices were used to assess the immune system state. Preventive application of WBC restored the number of lymphocytes and eosinophils and increased the number of monocytes in rats of both age groups, as well as the total leukocytes count in young animals. In animals of both age groups, young forms of neutrophils predominated, immunoreactivity decreased, the effector link of immunity and the macrophage system were activated. Delayed-type hypersensitivity processes in young rats as well as immediate-type hypersensitivity processes in aged rats were activated, and the reactive response of neutrophils decreased/increased according to the age of the animals.

Probl Cryobiol Cryomed 2024; 34(3):201–212

Dynamics of Ocular Surface Temperature in Rabbits During Transpalpebral Cooling (10°C)

Oleksandra Dorokhova — 2024-09-30

The study was aimed at evaluating the dynamics of the ocular surface temperature of healthy rabbits (chinchilla breed, 10 months’ age) in the ciliary body projection during artificial transpalpebral cooling (10°C). The study was conducted in 24 rabbits (48 eyes). Developed original thermoelectric devices were used for thermometry of the ocular surface and its cooling. The initial epibulbar temperature was measured by the contact method in the ciliary body projection prior to cooling. The eye was cooled by contact through closed eyelids with a constant temperature of the cooling surface of 10°C. Local epibulbar temperature was measured during 30 minutes of cooling every 10 minutes, as well as after stopping the cold exposure for 40 minutes. The decrease in the ocular surface temperature of rabbits in the ciliary body projection during the first 10 minutes was 4.6°C and for the next 20 minutes it was only 0.7°C. A rapid recovery of epibulbar temperature by 3.9°C was observed within the first 10 minutes after cessation of cold exposure, followed by delayed warming of the ocular surface by only 0.6°C for the next 30 minutes. Transpalpebral contact controlled ocular cooling at 10°C enabled to achieve a potentially therapeutic level of hypothermia of the structures of the ocular anterior segment, which led to the prospect of using the beneficial properties of artificial local hypothermia in ophthalmology.

Probl Cryobiol Cryomed 2024; 34(3):213–225

Cryolysate of Tumor Cells: From Technology of Its Obtaining up to Creation of Anti-сancer Vaccines

Anatoliy Goltsev — 2024-09-30

The possibility of obtaining immune dendritic cells (iDCs) from bone marrow mononuclear cells has been proven by adding granulocyte-macrophage colony-stimulating factor, interleukin-4, and tumor cell cryolysate to the culture as an inducer of their maturation. The source of tumor antigens for obtaining iDCs was a cryolysate of Ehrlich adenocarcinoma cells, which was obtained by 5-fold freeze-thawing. It was established that the differentiation of mononuclear cells into iDCs is accompanied by the loss of the CD14 antigen, the appearance of maturity marker characteristics of these cells (CD83, CD11c antigens) and an increase in the expression of costimulatory molecules (CD80, CD86). The research results enable not only to evaluate the qualitative characteristics of iDCs obtained thereby, but also to outline the prospects of their use in immune correction of oncopathology.

Probl Cryobiol Cryomed 2024; 34(3):226–235

Clinical Outcomes of Infertility Treatment Cycles for Repeatedly Cryopreserved Embryos

Maryna Petrushko — 2024-09-30

The study analyzed the survival, re-expansion, and implantation rates of human preimplantation embryos at the blastocyst stage following one and two rounds of cryopreservation using vitrification. Embryos subjected to repeated cryopreservation showed significantly lower survival, re-expansion, and implantation rates compared to those cryopreserved once (р < 0.05 for survival, р < 0.001 for re-expansion and implantation). The highest implantation rate was observed in the group with embryos cryopreserved once (р < 0.001). The results indicate that repeated cryopreservation negatively affects the viability and implantation of embryos, potentially due to structural and functional impairments. These findings highlight the importance of carefully assessing and considering potential risks when planning infertility treatment using repeatedly cryopreserved embryos.

Probl Cryobiol Cryomed 2024; 34(3): 236–244

Feasibility of Cord Blood Application in Cell Therapy for Central Nervous System Pathologies

Ivan Shcheniavskyi — 2024-09-30

This paper presents a search and analysis of scientific papers reporting about the findings aimed to elucidate the possibility of using cord blood (CВ) for therapy of nervous system pathologies. The authors found neither clinical protocols nor recommendations that have the valid or official status, etc. But a significant number of clinical trial protocols for treating neurological diseases with CB cell transplantation have been published. The review shows the results of some experimental studies in the models of central nervous system pathologies and clinical trials that have studied the CB therapeutic potential. The analysis of these reports suggests the prospects for using CВ in therapy of various pathologies, including neurodegenerative ones. However, further profound, targeted and well-funded research and extended clinical trials are needed to realize the therapeutic potential of CВ. The authors believe that in the nearest future it will be essential to study a paracrine effect of transplanted stem cells as an alternative to cell therapy.

Probl Cryobiol Cryomed 2024; 34(3):165–185