Influence of Sucrose on the Efficiency of Cryopreservation of Human Umbilical Cord-Derived Multipotent Stromal Cells with the Use of Various Penetrating Cryoprotectants

We studied the influence of sucrose applied in combination with different concentrations of penetrating cryoprotectants (DMSO, ethylene glycol, and glycerol) on the efficiency of cryopreservation of umbilical cord-derived multipotent stromal cells (MSC). The results indicate that these cells can be cryopreserved with the use of 5-10% DMSO or ethylene glycol with equal efficiency; addition of 0.2 M sucrose does not affect cell survival after thawing. The efficiency of glycerol as a cryoprotectant increases with increasing its concentration from 5 to 10%, but remains significantly lower than the efficiency of DMSO or ethylene glycol. Addition of sucrose to a final concentration of 0.2 M increases the efficiency of glycerol. The efficiency of combination of 10% glycerol and sucrose was comparable with that of combinations of DMSO and ethylene glycol with sucrose. The mechanism of the observed enhancement is apparently related to the influence of sucrose on the dynamic properties of the lipid membranes and facilitation of glycerol diffusion into the cells. © 2021, Springer Science+Business Media, LLC, part of Springer Nature

ВЛИЯНИЕ САХАРОЗЫ НА ЭФФЕКТИВНОСТЬ КРИОКОНСЕРВАЦИИ МУЛЬТИПОТЕНТНЫХ СТРОМАЛЬНЫХ КЛЕТОК ПУПОЧНОГО КАНАТИКА ЧЕЛОВЕКА С ИСПОЛЬЗОВАНИЕМ РАЗЛИЧНЫХ ПРОНИКАЮЩИХ КРИОПРОТЕКТОРОВ

Исследовали влияние сахарозы в сочетании с различными концентрациями проникающих криопротекторов (ДМСО, этиленгликоля и глицерола) на эффективность криоконсервации мультипотентных стромальных клеток (МСК) пупочного канатика. МСК могут быть успешно криоконсервированы с использованием ДМСО или этиленгликоля с равной эффективностью при содержании их в криорастворе в пределах 5-10%, причём добавление 0.2 М сахарозы не влияет на выживаемость клеток после размораживания. Эффективность глицерола в качестве криопротектора повышается с увеличением концентрации от 5 до 10%, однако остаётся значимо ниже, чем эффективность ДМСО и этиленгликоля. Добавление 0.2 М сахарозы повышает эффективность глицерола, при этом комбинация 10% глицерола и сахарозы имеет сравнимую с аналогичными комбинациями ДМСО и этиленгликоля эффективность. Предполагаемый механизм такого влияния заключается в воздействии сахарозы на динамические показатели липидной мембраны, которое обеспечивает более высокую скорость диффузии глицерола в клетку

КРИОКОНСЕРВАЦИЯ ТКАНИ ПУПОВИНЫ ЧЕЛОВЕКА: ПЕРСПЕКТИВЫ КЛИНИЧЕСКОГО ПРИМЕНЕНИЯ

The paper analyzes the data available in the modern scientific literature on the possibility of human umbilical cord tissue cryopreservation and its further clinical application. It describes protocols to obtain various biomedical products (vascular matrix, Wharton jelly-based grafts, and multipotent stromal cells) from the frozen umbilical cord, as well as prospects of their clinical use. The analysis of the literature data may be indicative of the active development of a promising cell biotechnology area, such as human umbilical cord cryopreservation. © Bionika Media Ltd.Проведен анализ имеющихся в современной научной литературе данных о возможности криоконсервации ткани пуповины человека и ее дальнейшем клиническом применении. Описаны протоколы для получения различных биомедицинских продуктов (матрикса сосудов, графтов на основе вартонова студня, мультипотентных стромальных клеток) из замороженной пуповины и перспективы их клинического применения. Проведенный анализ литературных данных позволяет предположить активное развитие такого перспективного направления клеточной биотехнологии как криоконсервация пуповины человека

Regenerative medicine of pancreatic islets

The pancreas became one of the first objects of regenerative medicine, since other possibilities of dealing with the pancreatic endocrine insufficiency were clearly exhausted. The number of people living with diabetes mellitus is currently approaching half a billion, hence the crucial relevance of new methods to stimulate regeneration of the insulin-secreting β-cells of the islets of Langerhans. Natural restrictions on the islet regeneration are very tight; nevertheless, the islets are capable of physiological regeneration via β-cell self-replication, direct differentiation of multipotent progenitor cells and spontaneous α- to β- or δ- to β-cell conversion (trans-differentiation). The existing preclinical models of β-cell dysfunction or ablation (induced surgically, chemically or genetically) have significantly expanded our understanding of reparative regeneration of the islets and possible ways of its stimulation. The ultimate goal, sufficient level of functional activity of β-cells or their substitutes can be achieved by two prospective broad strategies: β-cell replacement and β-cell regeneration. The "regeneration"strategy aims to maintain a preserved population of β-cells through in situ exposure to biologically active substances that improve β-cell survival, replication and insulin secretion, or to evoke the intrinsic adaptive mechanisms triggering the spontaneous non-β- to β-cell conversion. The "replacement"strategy implies transplantation of β-cells (as non-disintegrated pancreatic material or isolated donor islets) or β-like cells obtained ex vivo from progenitors or mature somatic cells (for example, hepatocytes or α-cells) under the action of small-molecule inducers or by genetic modification. We believe that the huge volume of experimental and clinical studies will finally allow a safe and effective solution to a seemingly simple goal-restoration of the functionally active β-cells, the innermost hope of millions of people globally. ©The Author(s) 2020

Chronology of quaternary volcanic activity in Keli highland (Greater Caucasus) on evidence of K-Ar isotope dating

The specified on the basis of K-Ar data magmatic activity phases in manifestation time coincide with certain activity phases of other large latest-volcanism centers at Greater Caucasus - Elbrus and Kazbek. The initial volcanic activity phase (225-175 thousand years ago) was marked by extrusion bodies formation in different parts of the region. In the Late neo-Pleistocene (130-60 t.y.a.) the activity had reached its maximum, at that stage predominantly lava volcanoes erupted and thus lavatic plateaus and valley streams were formed. The youngest geologic formations in the western part of Keli highland are the volcano Sev. Shadilkhokh and associated with it Hodzh stream; however, strict proof of their Holocene age requires additional data

Chronology of quaternary volcanic activity in Keli highland (Greater Caucasus) on evidence of K-Ar isotope dating

The specified on the basis of K-Ar data magmatic activity phases in manifestation time coincide with certain activity phases of other large latest-volcanism centers at Greater Caucasus - Elbrus and Kazbek. The initial volcanic activity phase (225-175 thousand years ago) was marked by extrusion bodies formation in different parts of the region. In the Late neo-Pleistocene (130-60 t.y.a.) the activity had reached its maximum, at that stage predominantly lava volcanoes erupted and thus lavatic plateaus and valley streams were formed. The youngest geologic formations in the western part of Keli highland are the volcano Sev. Shadilkhokh and associated with it Hodzh stream; however, strict proof of their Holocene age requires additional data

ТКАНЕИНЖЕНЕРНАЯ КОНСТРУКЦИЯ НА ОСНОВЕ ПОЛИДИОКСАНОНА И МУЛЬТИПОТЕНТНЫХ СТРОМАЛЬНЫХ КЛЕТОК ДЛЯ ПЛАСТИКИ ДЕФЕКТОВ БРЮШНОЙ ПОЛОСТИ И ДНА МАЛОГО ТАЗА

Objective. To study the safety and efficiency of transplantation of the developed tissue-engineered prosthesis based on polydioxanone and cultured multipotent stromal cells (MSC) from the umbilical cord. Material and methods. In vivo biocompatibility of prostheses was evaluated in outbred rats, by modeling a full-layer defect of the anterior abdominal wall, the edges of which was sutured with a polydioxanone prosthesis that was unpopulated and populated by the cultured cells. A prosthesis based on the decellularized dermis (Permacol) was used in the comparison group. The animals were withdrawn from the experiment at 3, 10, 30, 60, and 180 days after surgery. Macroscopic, tensiometric, histomorphometric, and immunohistochemical studies were conducted. Results. The polydioxanone-based prostheses were found to be more effectively integrated and grow their own tissues. The biomechanical properties of tissues in the field of transplantation in the long-term periods did not differ between the groups and the native tissue of the anterior abdominal wall. When a tissue-engineered construct was transplanted, there was a lower inflammatory response due to macrophage M2 polarization, as well as a more pronounced angiogenesis. The transplanted cells did not differentiate into blood vessel cells and were totally eliminated by a recipient’s macrophages. The registered effects appeared to be shown via paracrine mechanisms. Conclusion. The addition of cultured MSCs to the prosthesis could substantially reduce the severity of an inflammatory response to rejection of a foreign body and stimulate angiogenesis and the rate of replacement by the recipient’s own tissues. © 2018, Bionika Media Ltd.. All rights reserved.Цель исследования. Изучение безопасности и эффективности трансплантации разработанного тканеинженерного протеза на основе полидиоксанона и культуры мультипотентных стромальных клеток (МСК) пупочного канатика. Материал и методы. Оценку биосовместимости протезов in vivo проводили на беспородных крысах путем моделирования полнослойного дефекта передней брюшной стенки, к краям которого подшивали полидиоксаноновый протез, незаселенный или заселенный культурой клеток. В качестве группы сравнения использовали протез на основе децеллюляризированной дермы (Permacol). Животных выводили из эксперимента на 3, 10, 30, 60 и 180 сутки после операции. Проводили макроскопическое, тензиометрическое, гисто-морфометрическое, иммуногистохимическое исследования. Результаты. Протезы на основе полидиоксанона более эффективно интегрировались и прорастали собственными тканями. Биомеханические свойства тканей в области трансплантации на отдаленных сроках не отличались между группами и нативной тканью передней брюшной стенки. При трансплантации тканеинженерной конструкции наблюдали меньшую степень выраженности воспалительной реакции за счет поляризации макрофагов в М2-направлении и более выраженный ангиогенез. Трансплантированные клетки не дифференцировались в клетки кровеносных сосудов и тотально элиминировались макрофагами реципиента. По-видимому, регистрируемые эффекты реализовались за счет паракринных механизмов. Заключение. Благодаря добавлению в состав протеза культуры МСК удалось существенно снизить степень выраженности воспалительной реакции отторжения инородного тела, стимулировать ангиогенез и скорость замещения собственными тканями реципиента

DMSO-Free Cryopreservation of Human Umbilical Cord Tissue

Human umbilical cord represents a source of multipotent stromal cells of a supreme therapeutic potential. The cells can be isolated from either fresh or cryopreserved umbilical cord tissues. DMSO is a cryoprotectant most commonly used for preservation of umbilical cord tissues; however, cyto- and genotoxicity of this compound is evident and well documented. In the present study we performed successful cryopreservation of the umbilical cord tissue using other cryoprotectants: propylene glycol, ethylene glycol, and glycerol. Of these, 1.5 M ethylene glycol and 20% glycerol turned out to be the best in terms of the preservation of living cells within the frozen tissue, early onset of migration of these cells out of the thawed explants, and overall efficacy of multipotent stromal cell isolation. Cryobanking of tissues can improve availability of multiple cell products for medical purposes and promote the development of personalized medicine. © 2018, Springer Science+Business Media, LLC, part of Springer Nature