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Dynamics of Dimethyl Sulfoxide Penetration Into L929 Cells and L929-Based Spheroids 二甲基亚砜对L929细胞和L929基球体的渗透动力学
Q4 Medicine Pub Date : 2021-12-25 DOI: 10.15407/cryo31.04.316
A. Moisieiev, I. Kovalenko, S.Ju. Kovalenko, G. Bozhok, O. Gordienko
The study proposes an algorithm for calculating of appreciable permeability coefficients for multicellular structures in a cryoprotectant medium using physical and mathematical model of mass transfer. The values of surface-area-to-volume ratio for L929 cells at different temperatures were determined and the thermal expansion coefficient of the surface area of cell membranes was calculated (β = 2.7 × 10-3 /°C). The osmotically inactive volume for L929 cells and their spheroids was determined. Filtration and permeability coefficients to DMSO for L929 cells and in toto spheroids were found from the dynamic curves of relative volume change. The calculated parameters are the highest for individual cells and significantly (p <0.05) decrease for cells in the spheroids with increasing depth of their location, this reduction may be stipulated by a decrease in the available surface of cells in the spheroids for the penetration of extracellular substances. Obtained in this research permeability characteristics of spheroids can be used to develop optimal cryopreservation regimens for them.
本研究提出了一种利用传质物理模型和数学模型计算低温保护剂介质中多细胞结构可观渗透系数的算法。测定了不同温度下L929细胞的表面积体积比,计算了细胞膜表面积的热膨胀系数(β = 2.7 × 10-3 /°C)。测定了L929细胞及其球体的渗透失活体积。通过相对体积变化的动态曲线,得到了L929细胞和toto球体对DMSO的过滤系数和渗透系数。计算出的参数在单个细胞中是最高的,而在球体中的细胞随着其位置深度的增加而显著(p <0.05)降低,这种降低可能是由于球体中细胞用于细胞外物质渗透的可用表面的减少所规定的。本研究获得的球类材料的渗透性特征可用于制定球类材料的最佳低温保存方案。
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引用次数: 2
Anti-Inflammatory Properties of Diclofenac Sodium on Background of Its Combined Use With Cryopreserved Placenta Extract in Experiment 双氯芬酸钠与胎盘冻存液联合应用的抗炎作用研究
Q4 Medicine Pub Date : 2021-12-25 DOI: 10.15407/cryo31.04.364
F. Hladkykh
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引用次数: 7
Placental Cryoextract Corrects the Level of Thyroid Hormones in Rats 胎盘冷冻提取物对大鼠甲状腺激素水平的影响
Q4 Medicine Pub Date : 2021-12-25 DOI: 10.15407/cryo31.04.353
N. Malova, I. Komarova, L. Sirotenko, N. Brechka, Yuliya Kurilko, T. Komisova, A. Goltsev
Effectiveness of cryopreserved placental drug ‘Placental cryoextract’ to correct the reproductive system disorders, occurring against the background of low levels of thyroid hormones, was studied in 50 male rats with mercazolyl hypothyroidism. Using standard test systems for animals, the level of total and free forms of thyroid hormones, total testosterone and estradiol was determined. The morphofunctional state of sperm (number, motility, pathological forms) was traditionally assessed. It was shown that the introduction of the biological product (at a dose of 50–150 μl / kg) restored the functional activity of the thyroid gland of animals, and the lowest dose was more effective. Placental cryoextract also had a dose-dependent effect on the reproductive system of animals with hypothyroidism: the most effective dose was 150 μl / kg, which maximally restored the morphofunctional parameters of sperm and stimulated testosterone production as well as normalized the Ts / E2 ratio. Cryopreserved placenta extract can be considered as a potential drug for the correction of hypothyroidism and reproductive disorders of endocrine origin in male rats.
在50只患有甲唑啉甲状腺功能减退症的雄性大鼠中,研究了冷冻保存的胎盘药物“胎盘冷冻提取物”在甲状腺激素水平低的背景下纠正生殖系统疾病的有效性。使用动物标准测试系统,测定甲状腺激素、总睾酮和雌二醇的总形式和游离形式的水平。精子的形态功能状态(数量、活力、病理形式)传统上是评估的。研究表明,生物制品的引入(剂量为50–150μl/kg)恢复了动物甲状腺的功能活性,最低剂量更有效。胎盘冷冻提取物对甲状腺功能减退动物的生殖系统也有剂量依赖性影响:最有效剂量为150μl/kg,最大限度地恢复了精子的形态功能参数,刺激了睾酮的产生,并使Ts/E2比值正常化。冷冻保存的胎盘提取物可以被认为是一种潜在的药物,用于纠正雄性大鼠甲状腺功能减退和内分泌生殖障碍。
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引用次数: 0
Cultural Properties of Cryopreserved Thymic Multipotent Stromal Cells and Fetal Skin and Muscle-Derived Cells 胸腺多能基质细胞及胎儿皮肤和肌肉来源细胞的培养特性
Q4 Medicine Pub Date : 2021-09-25 DOI: 10.15407/cryo31.03.249
Valentina Nikolska, Y. Semenova, Lyuba Taranukha, Ihor Nikolsky
The paper provides a comparison of properties of cryopreserved fetal murine multipotent stromal cells (MSCs) of skin-muscular origin and those derived from adult thymus in culture in vitro. Fetal MSCs showed a 30% higher number of average population doublings within 24 hrs, and 41% lower average population doubling time. It was found that the fetal MSCs of the 4th passage had a 39% higher clonogenic activity than the adult thymus-derived ones. Fetal MSCs and those derived from adult thymus differentiated in osteogenic and adipogenic lineages with equal efficiency in special culture media. Fetal and thymus-derived MSCs were characterized by almost the same high ability of contact interaction with thymocytes, and the fibroblast-lymphocyte rosette (FLR) formation. They were far less active in FLR formation with lymph node cells. This indicated the presence of membrane affinity for immature lymphoid cells in both MSC subpopulations. The results showed the fetal MSCs to be significantly different from the adult thymus-derived MSCs by more active kinetics of growth and clonogenic potential. However, both cell subpopulations had virtually the same ability for linear differentiation and showed high activity during contact with immature lymphoid cells. Linear differentiation and the ability to interact with lymphocytes were found to be quite stable properties of MSCs, but a proliferative activity and in vitro colony formation distinguished significantly in different types of MSCs. This can be taken into account when choosing the cells for therapy, research and results assessment.
本文比较了冷冻保存的皮肤肌肉来源的胎鼠多能基质细胞(MSCs)和来源于成人胸腺的MSCs在体外培养中的特性。胎儿间充质干细胞在24小时内平均群体倍增次数增加30%,平均群体倍增时间减少41%。研究发现,第4代的胎儿MSCs的克隆活性比成人胸腺来源的MSCs高39%。胎儿MSCs和来自成人胸腺的MSCs在成骨和成脂谱系中在特殊培养基中以相同的效率分化。胎儿和胸腺来源的骨髓间充质干细胞具有几乎相同的与胸腺细胞接触相互作用的高能力,以及成纤维细胞淋巴细胞花环(FLR)的形成。它们在与淋巴结细胞形成FLR中的活性要低得多。这表明在两个MSC亚群中都存在对未成熟淋巴细胞的膜亲和力。结果表明,胎儿MSCs与成人胸腺来源的MSCs在生长动力学和克隆潜能方面有显著差异。然而,两种细胞亚群几乎具有相同的线性分化能力,并且在与未成熟淋巴细胞接触时表现出高活性。线性分化和与淋巴细胞相互作用的能力被发现是MSCs相当稳定的特性,但不同类型的MSCs的增殖活性和体外集落形成显著不同。在选择用于治疗、研究和结果评估的细胞时,可以考虑到这一点。
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引用次数: 0
Theoretical Estimation of Optimal Linear Cooling Rate for PK-15 Cell Suspension PK-15细胞悬浮液最佳线性冷却速率的理论估计
Q4 Medicine Pub Date : 2021-09-25 DOI: 10.15407/cryo31.03.214
O. Gordienko, I. Kovalenko, S.Ju. Kovalenko, L. Kuleshova, O. Todrin
Preservation of cells during crystallization of the cell suspension is influenced by two types of damaging factors. The first type of cryoinjury occurs during the crystallization of the extracellular environment and is caused by dehydration of cells, increasing the concentration and ionic strength of extracellular and intracellular solutions. As the cooling rate rises, the damage rate of the first type decreases as a result of the reduced time of action of damaging factors. The second type of cryoinjury is intracellular crystallization, the probability of which enhances at high cooling rates, is considered the most destructive to cells. The optimal linear cooling rate for PK-15 cells is determined using a physico-mathematical model, which describes the probability of cryoinjury of cells in the linear freezing mode and is based on the two-factor theory of cryoinjury, thermodynamic theory of homogeneous crystallization and general theory of activation-type processes. The findings have shown that within the range of cooling rates < 0.5 °C/min the cryoinjury of PK-15 cells occurs mainly due to the effects of the solution, and at cooling rates > 2.5 °C/min this was mainly resulted from an intracellular crystallization. The dependence of the percentage of damaged cells on the cooling rate has a relatively wide minimum within the range of cooling rates of 0.5 °C/min… 2.5 °C/min.
细胞悬浮液结晶过程中细胞的保存受到两类破坏因素的影响。第一种类型的低温损伤发生在细胞外环境的结晶过程中,是由细胞脱水引起的,细胞外和细胞内溶液的浓度和离子强度增加。随着冷却速率的增大,由于损伤因素作用时间的缩短,第一类损伤速率降低。第二种类型的低温损伤是细胞内结晶,其可能性在高冷却速率下增强,被认为是对细胞最具破坏性的。基于冷冻损伤双因素理论、均匀结晶热力学理论和活化型过程的一般理论,利用描述细胞在线性冷冻模式下冷冻损伤概率的物理数学模型确定了PK-15细胞的最佳线性冷却速率。结果表明,在冷却速率< 0.5°C/min范围内,PK-15细胞的低温损伤主要是由于溶液的作用,而在冷却速率> ~ 2.5°C/min范围内,低温损伤主要是由于细胞内结晶引起的。在0.5°C/min ~ 2.5°C/min的冷却速率范围内,损坏电池的百分比对冷却速率的依赖性相对较小。
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引用次数: 1
Impact of Dorsal Root Ganglia Cryoextract on Histological Steatures in Dices and Contractility of Uterus in Differently Aged Rats 背根神经节冷冻提取物对不同年龄大鼠骰子组织学稳定性和子宫收缩能力的影响
Q4 Medicine Pub Date : 2021-09-25 DOI: 10.15407/cryo31.03.258
H. Nesteruk, V. Ustichenko, N. Alabedalkarim, V. Padalko, O. Protsenko, Ievgen Legach
To date, the number of women giving birth to their first child in late reproductive age is increasing around the world. This stipulates a need in designing the new approaches to restore the uterine contractile activity. In this paper, the histological features of uterus and its contractile activity have been experimentally studied in differently aged rats after the dorsal root ganglia cryoextract (DRGCE) administration. The cryoextract was derived from dorsal root ganglia of neonatal piglets by three-fold freezing down to –196°C in saline, followed by thawing at room temperature, homogenization and centrifugation. Here, we used the female rats of reproductive age (RA, 6-month-old) and those of late reproductive age (LRA, 14-month-old). Animals of both age groups received intraperitoneally either DRGCE (experimental groups) or saline (control groups) for 9 days by 0.2 ml. To days 28–29 after administration beginning the animals were sacrificed and the uterine fragments were taken for histological examination and study of oxytocin (OT)-induced uterine contractile activity (UCA). The strength of OT-induced uterine isometric contraction was found to decrease by 28.6% in LRA rats vs. the RA ones. The DRGCE administration to LRA rats increased the UCA indices, namely the contractile frequency, contractile amplitude and strength of isometric contraction augmented by 25, 9.8 and 30% respectively, as compared with the group of the same age without DRGCE introduction. This effect was observed on the background normal uterine histological structure and unchanged myometrial thickness.
迄今为止,在世界范围内,晚育妇女生育第一胎的人数正在增加。这就要求设计恢复子宫收缩活动的新方法。本文通过实验研究了不同年龄大鼠背根神经节冷冻提取液(DRGCE)后子宫的组织学特征及其收缩活动。从新生仔猪背根神经节中提取冷冻提取物,在生理盐水中三次冷冻至-196°C,然后在室温下解冻,均质和离心。本实验选用育龄雌性大鼠(RA, 6月龄)和晚期育龄雌性大鼠(LRA, 14月龄)。两组动物分别腹腔注射DRGCE(实验组)或生理盐水(对照组)0.2 ml,持续9天。给药后28-29天,处死动物,取子宫碎片进行组织学检查和催产素(OT)诱导的子宫收缩活动(UCA)研究。ot诱导的子宫等长收缩强度比RA大鼠降低28.6%。与未加DRGCE组相比,DRGCE使LRA大鼠的UCA指数(即收缩频率、收缩幅度和等距收缩强度)分别提高了25%、9.8和30%。这种效果是在子宫组织结构正常、子宫肌层厚度不变的背景下观察到的。
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引用次数: 1
Influence of Ways to Thaw Bull Sperm on Its Quality 牛精子解冻方式对其品质的影响
Q4 Medicine Pub Date : 2021-09-25 DOI: 10.15407/cryo31.03.277
Yu. A. Shakhova, A. Paliy, A. Paliy, O. Shkromada, Y. Musiienko, I. Bondarenko
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引用次数: 2
Role of Liquid-Liquid Phase Transitions in Mechanism of Erythrocyte Protection During Cooling with CRIHBT-115 Cryopreservative Agent 液-液相变在CRIHBT-115抗冻剂冷却过程中红细胞保护机制中的作用
Q4 Medicine Pub Date : 2021-09-25 DOI: 10.15407/cryo31.03.236
O. Khodko
The presence in the system of critical liquid-liquid phase transition (PT) by the mechanism, resulting in formation of dispersion system, namely high-concentrated emulsion, has been established here during cooling when using polarized light microscopy and fixation of critical opalescence phenomenon in erythrocyte concentrate with glycerol-containing cryopreservative agent, designed at the Central Research Institute of Haematology and Blood Transfusion (Russia) (CRIHBT-115 ). The studied cryobiological system displayed no signs of crystallization. A phase behaviour of cryopreservative and supernatant has been studied during cooling-warming cycle. Changes in the volume of cryopreservative and erythrocyte concentrate were comparatively and qualitatively evaluated during cooling. The mechanism of protective action of cryopreservation solution has been determined. The similarity between physical and chemical processes during cooling-warming of erythrocyte cytoplasm and garlic meristem cells (germinal plant tissue) when entering cold anabiosis has been established.
在俄罗斯中央血液学和输血研究所(CRIHBT-115)设计的含甘油冷冻保鲜剂对红细胞浓缩物的临界乳光现象进行偏光显微镜和固定的冷却过程中,建立了临界液-液相变(PT)体系中存在的机制,从而形成分散体系,即高浓度乳化液。所研究的低温生物系统没有显示出结晶的迹象。研究了低温保鲜剂和上清液在冷暖循环中的相行为。在冷却过程中,对冷冻保鲜剂和红细胞浓缩物的体积变化进行了比较和定性评价。确定了低温保存液的保护作用机制。红细胞细胞质和大蒜分生组织细胞(植物生发组织)进入低温复苏时的冷暖物理和化学过程具有相似性。
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引用次数: 0
Manufacturing Characteristics of Probiotic Esherichia Coli Strain M-17 After Immobilization in Gel Carriers and Storage at Various Low Temperatures 大肠埃希氏菌M-17凝胶载体固定化及不同低温贮藏后的生产特性
Q4 Medicine Pub Date : 2021-09-25 DOI: 10.15407/cryo31.03.273
I. Petrov, I. Vysekantsev, Y. Cherkashina, O. Peretyatko, Yu. Yagniuk
Іммобілізовані пробіотики все більше застосовують у виробництві пробіотичних препаратів, лікувально-профілактичних продуктів харчування, кормових домішок [6]. Технології довгострокового зберігання комерційних препаратів і продуктів на основі іммобілізованих пробіотиків знаходяться в стадії розробки. Найбільш ефективними і широко вживаними методами консервування вільних клітин різних мікроорганізмів є ліофілізація і зберігання за низьких температур [5]. Мета роботи – вивчення збереженості виробничих характеристик пробіотичного штаму Esherichia coli M-17 (E. coli M-17) після іммобілізації в гелевих носіях і зберігання за різних низьких температур.
固定化抗生素越来越多地用于抗生素生产、医学预防食品、食品之家[6]。商业产品和基于固定蛋白的产品的长期储存技术正处于开发阶段。保存不同微生物的游离细胞最有效和最广泛使用的方法是冷冻干燥和低温储存[5]。本工作的目的是研究在凝胶介质中动员和在不同低温下储存后,大肠杆菌M-17(E.coli M-17)的亲生物特性的保守性。
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引用次数: 0
Thermoplastic Analysis of Cluster Crystallization of Cryoprotective Solutions 低温保护溶液团簇结晶的热塑性分析
Q4 Medicine Pub Date : 2021-09-25 DOI: 10.15407/cryo31.03.203
Oleksandr Osetskyi, T. Gurina, Anna Poliakova, Stanislav Sevastianov
For the first time the possibility of using thermomechanical analysis to construct the fragments of state diagrams of cryoprotective solutions in the zone of glass transition temperatures has been considered. A method for studying cluster crystallization of cryoprotective solutions based on thermomechanical curves has been developed. The parameters of thermomechanical curves of frozen aqueous solutions of dimethyl sulfoxide (DMSO), polyethylene oxide 1500 (PEO-1500), glycerol were analyzed and the relationship between these parameters and the cluster crystallization kinetics for these solutions was established. On the basis of experimentally obtained thermomechanical curves for the frozen solutions of DMSO and PEO-1500 the possibility of formation of clusters of two types has been shown: on the basis of ice and cryoprotective substance microcrystals. Additional experimental data were obtained to construct a complete state diagrams of cryoprotective solutions, which include the existing cluster phase areas.
首次考虑了在玻璃化转变温度区使用热机械分析构建冷冻保护溶液状态图片段的可能性。开发了一种基于热机械曲线研究低温保护溶液簇结晶的方法。分析了二甲基亚砜(DMSO)、聚环氧乙烷1500(PEO-1500)、甘油冷冻水溶液的热力学曲线参数,建立了这些参数与溶液团簇结晶动力学的关系。根据实验获得的DMSO和PEO-1500冷冻溶液的热机械曲线,已经表明了形成两种类型团簇的可能性:基于冰和冷冻保护物质微晶。获得了额外的实验数据,以构建冷冻保护溶液的完整状态图,其中包括现有的簇相区域。
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引用次数: 1
期刊
Problems of Cryobiology and Cryomedicine
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