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The review considers the possibility of using modern nanotechnological developments aimed to achieve alternative cryobiological goals. On the one hand, the use of nanomaterials will increase the functional value of thawed cells due to such unique characteristics of nanoparticles as size, shape, surface charge, chemical composition, etc. Nanomaterials can be used as nanocontainers for impermeable cryoprotective agents (CPAs) and cause significant changes in crystal formation, thermal conductivity and other properties of cells, tissues and organs, that increases the efficiency of their cryopreservation. On the other hand, the combined use of nanomaterials and low-temperature freezing factors is considered a promising method of destruction of pathologically altered cells and tissues, as it minimizes the risk of recurrence of oncopathology after insufficient freezing-out of the tumor site. Probl Cryobiol Cryomed 2020; 30(4): 313–330

Goltsev, A., Fuller, B., Bondarovich, M., Babenko, N., Gaevska, Y., Buriak, I., Dubrava, T. ., Yampolska, K., Lutsenko, O., & Ostankov, M. (2020). COVID–19 – потенційна мішень для кріобіології і кріомедицини. Проблеми кріобіології і кріомедицини, 30(2), 107–131. https://doi.org/10.15407/cryo30.02.107 У огляді представлено дані щодо імунопатогенезу COVІD-19 і підходів до його профілактики та лікування. Низька ефективність противірусних засобів обумовлена здатністю вірусу SARS-CoV2 до зміни власних структурних і функціональних характеристик. Існуючі стратегії лікування COVІD-19 сфокусовані на застосуванні лікувальних засобів прямої противірусної дії, модуляції вродженої імунної відповіді, пригніченні «цитокінового шторму» та застосуванні плазми реконвалесцентів. Дизрегуляція взаємодії систем вродженого та адаптивного імунітету обумовлює запуск аутоімунного процесу в організмі вірусоносія, що потребує застосування альтернативних підходів до профілактики таких захворювань із використанням кріобіологічних технологій. Наведено результати вивчення імунобіологічної активності кріоконсервованого лейкоконцентрату кордової крові людини (кЛККЛ) і його компонентів за превентивного інтраназального введення. Ефективність застосування кЛККЛ пов’язана з можливістю перепрограмування генів, відповідальних за реалізацію імунних відповідей організму та індукцію «тренованого» імунітету. Така модифікація стану імунної системи може бути найбільш перспективною у забезпеченні захисту організму від вірусів, зокрема COVІD-19. __________________________________________________________________________________________ The review presents data on immune pathogenesis of COVID-19 and approaches to its prevention and treatment. The low effectiveness of antiviral drugs is caused by the ability of SARS-CoV2 virus to change its own structure and functions. Existing treatment strategies of COVID-19 are focused on applying the direct antiviral therapies, modulation of innate immune response, "cytokine storm" suppression and use of convalescent plasma. Deregulated interaction between innate and adaptive immune systems determines the start of autoimmune process in the virus carrier body, that requires the use of alternative ways to prevent such diseases using cryobiological technologies. The results of studying the immune biological activity of cryopreserved human cord blood leukoconcentrate (cHCBL) and its components by preventive intranasal administration have been presented. The effectiveness of cHCBL is related to the possible reprogramming of the genes responsible for the implementation of the body's immune responses and induction of the "trained" immunity. Such a modification of the immune system state can be the most promising in protecting the body against viruses, in particular COVID-19.

This paper presents the investigations on the impact of lyophilisation conditions on structural and functional characteristics of human cord blood leukoconcentrate (HCBL) such as: the number and viability, cytomorphological indices, content of hematopoietic stem (CD34+) and T-regulatory cells (CD4+CD25high), hematopoietic colony forming units in culture (CFUc). Four regimens, including two programs with and without 3.5% DMSO were used for hcbl cell lyophilisation. The programs dii¬€ered in temperature value and duration wherein the primary drying proceeded. When using the i¬rst program, the cells were dried for 10 hrs (–28ÂoÐi), and 9 hrs (–18ÂoÐi) for the second one. The designed method for HCBL lyophilisation, suggested the use of the DMSO-free i¬rst program, ensured the cell preservation in a heterogeneous population of cord blood according to all the studied characteristics. Probl Cryobiol Cryomed 2019; 29(4): 332-343

Cryoablation is a method of choice in treatment of solid tumors of dii¬€erent localization. In addition to the destruction of pathological tissue, the cryoablation causes to complex changes in the immune system. This paper briei¬‚y discusses the implementation of the abscopic ei¬€ect after cryosurgery of tumors, as well as possible changes in the immune system that accompany it. The abscopic ei¬€ect is achieved with a release of antigens from the destroyed cells, which trigger a cascade of immune reactions. This article summarizes the results of the combined use of cryoablation and immunotherapeutic methods, in particular adoptive therapy. The introduction of dendritic cells in combination with cryoablation can signii¬cantly enhance antitumor immunity and reduces the risk of relapse. Probl Cryobiol Cryomed 2019; 29(4): 297-302

Goltsev AM, Bondarovych MO, Babenko NM, Gaevska YO, Dubrava TG, Ostankov MV. Effect of different cryopreservation regimens on Ehrlich carcinoma growth. Cell Tissue Bank. 2019 Sep;20(3):411-421. doi: 10.1007/s10561-019-09780-9. Epub 2019 Jun 25. PMID: 31240496. The freezing rate is a decisive factor in determining the purpose of using low temperatures, i.e., for cryoablation or cryobanking of tumor cells. The research aim was to determine effect of different cryopreservation regimens on Ehrlich carcinoma (EC) growth in vivo and subpopulation composition of the formed ascites. The previously cryopreserved with slow and rapid rates EC cells were cultured in peritoneal cavity (PC) of mice for 7 days. Absolute number of cells in the PC, the subpopulation composition of tumor with flow cytometry using CD44 and CD24 markers were determined. Immediately after warming, a significant redistribution of EC subpopulation composition with a decreased content of the most tumorigenic CD44high cells after both freezing regimens was found. Culturing in vivo for 7 days contributed to the restoration of EC subpopulation composition, but with some a decrease in the tumor growth intensity when slow cooling was used. Rapid cooling contributed to significant inhibition of tumor growth with a reduced number of CD44+ and increased CD24+ cells. None of the cryopreservation regimens resulted in a complete elimination of tumorigenic CD44high tumor cells. The freezing rate determines the preservation of the subpopulation composition of the EC and intensity of its growth in vivo.

The review highlights some areas of malignant tumor treatment, particularly the cryosurgery of solid tumors. There is discussed the possibility of using the method of cryoablation as a tool for targeting and inactivation of cancer stem cells, being the main structural unit, initiating, maintaining growth and tumor metastasis. The publications regarding the phenotypic and functional characteristics of tumor-inducing cells, including both more potent CSCs and advanced in the differentiation of their descendants, the nature of cold factor impact on them were analyzed. The main ways to optimize the use of cryoablation technique in clinical practice are under consideration. Probl Cryobiol Cryomed 2015; 25(3):205-218

One of the tasks of current oncology is identification of cancer stem cells and search of therapeutic means capable of their specific inhibition. The paper presents the data on phenotype characteristics of Ehrlich carcinoma cells as convenient and easy-to-follow model of tumor growth. The evidence of cancer stem cells as a part of Ehrlich carcinoma and significance of CD44+ and CD44– subpopulations in maintaining the growth of this type of tumor were demonstrated. A high (tenfold) tumorigenic activity of the Ehrlich carcinoma CD44+ cells if compared to CD44– cells was proven. In this pair of comparison, the CD44+ cells had a higher potential of generating in peritoneal cavity of CD44high, CD44+CD24–, CD44+CD24+ cell subpopulations, highlighting the presence of cancer stem cells in a pool of CD44+ cells. In this study, the ability of synthesized hybrid nanocomplexes, comprising the nanoparticles of rare earth orthovanadates GdYVO4:Eu3+ and cholesterol to inhibit the tumor growth and to increase the survival of the animals with tumors was established. A special contribution into tumor-inhibiting effect is made by each of its components. Treatment of Ehrlich carcinoma cells with two-component hybrid complex resulted in maximum reduction in the concentration of the most tumorigenic CD44high cells with simultaneous rise in the number of CD117+ cells that decreased an intensity of tumor growth by 74.70 ± 4.38% if compared with the control.

Rheumatoid arthritis (RA) is one of the most widespread chronic inflammatory diseases of the joints with multifactor etiology. A crucial point in RA is the disorder of proliferation and apoptosis in immunocompetent cells as well as in the cells of monocyte-phagocyte system (MPS). Using the model of adjuvant arthritis (AA) in animals we comparatively assessed the development of apoptotic processes in MPS cells before and after administration of native and cryopreserved fetal liver cells (cFLCs). By means of Western blot analysis we showed the peculiarities of CD-95 cascade proteins expression in MPS cells during development of AA. After administration of cFLCs we found more pronounced proapoptotic effect, which degree depended on the period of their administration. Probl Cryobiol Cryomed 2014; 24(3):249-261.

The performed comparative analysis of structural and functional status of fetal liver (FL) hematopoietic stem cells (HSCs) of 14 gestation days and adult bone marrow (BM) of 4-month-old CBA/H mice demonstrated not only the differences of their initial qualitative and quantitative characteristics but also different responses to the effect of cryopreservation factors. Either FL or BM cells were frozen according to various regimens under protection of 5 and 10% dimethyl sulfoxide. To estimate structural and functional characteristics of HSCs derived from different sources there were used cytofluorimetry (content of CD34+СD38– cells), methods of colony formation in vivo (CFUs) and in vitro (CFU-GM). It has been established that certain cryopreservation regimens are able to manifest the effect of selective enrichment of cell populations with phenotype of hemopoietic progenitors (CD34+CD38– cells). Comparing the content of CFUs, CFU-GM, HSCs and cells with CD34+CD38– phenotype has shown that cryopreservation regimen for FL cells was not optimal for BM. It has been demonstrated that varying cryopreservation conditions allowed not only to provide optimal preservation rate of hemopoietic progenitors derived from FL and BM with various initial functional status, but to control it purposefully. Probl Cryobiol Cryomed 2014; 24(2):118-131.

The study was conducted to assess the effect of gestation term and cryopreservation factors on some functional parameters of mesenchymal stem cells (MSCs) in fetal liver (FL), in particular, ido gene expression rate. Expression rate of ido gene was decreased between 14th to 18th days of FL gestation. Following cryopreservation the expression rate of ido gene in FL cells of 18th gestation day increased significantly, and this level exceeded significantly the one in non-frozen samples of the 14th and 18th gestation day. This fact emphasizes the ability of cryopreservation to implement the ‘revitalizing’ effect in respect of FL cells of late gestation terms. Thus, cryopreservation can be used as the method of specific enhancing of MSCs immune modulating activity. Probl Cryobiol Cryomed 2014; 24(1):16–27.