Выращивание Ipomoea batatas (L.) Lam. в условиях светокультуры in vitro и ex vitro

Relevance. Currently, food products that include prebiotics, in particular, inulin, are particularly popular. Interest in this substance is justified by its valuable properties – it is a good immunomodulator, cleanses the body of toxins, radionuclides, "bad" cholesterol, promotes the assimilation of useful trace elements necessary for human life. Inulin is contained in plants such as jerusalem artichoke, chicory, as well as in sweet potatoes, the popularity of which is increasing every year. However, sweet potato plants are afraid of cold and frost-resistant. Therefore, the creation of new varieties and hybrids that are resistant to low temperatures is an urgent problem. Cellular biotechnology is aimed at solving this problem using methods of clonal microreproduction, cell selection, somatic hybridization, etc. For rapid reproduction and obtaining high-quality planting material, biotechnology methods are used, in particular, clonal micro-propagation. However, in this technology there are difficulties associated with poor adaptation of microclones to ex vitro conditions. This fact introduces an additional requirement for the selection of optimal rooting modes in vitro and ex vitro adaptation of microclones.Material and methodology. The aim of the work was to study the influence of cultivation conditions on in vitro rooting and ex vitro adaptation of I. batatas (L.) microclones. The object of the study was sweet potato microgears propagated in vitro. I. batatas micro-gears were cultured in vitro on a Murashige-Skug medium, differing by the type of auxins. The influence of red (R) and far red (FR) light on the shoots rooting in vitro and the adaptation of microclones ex vitro was studied.Results. It has been experimentally established that the cultivation of micro-gears on a medium containing indolyl butyric acid at a concentration of 0.5-1 mg/l and under conditions of illumination by LED lamps of red and far red light in equal amounts leads to the production of microclones with a well-developed root system and vegetative biomass. The use of an aeroponic installation at the last stage of clonal micro-propagation makes it possible to obtain high-quality planting material that can adapt well to open ground conditions.Актуальность. В настоящее время особой популярностью пользуются продукты питания, в состав которых входят пребиотики, в частности, инулин. Интерес к данному веществу оправдан его ценными свойствами – он является хорошим иммуномодулятором, очищает организм от токсинов, радионуклидов, «плохого» холестерина, способствует усвоению полезных микроэлементов, необходимых для жизнедеятельности человека. Инулин содержится в таких растениях, как топинамбур, цикорий, а также в батате, популярность которого с каждым годом возрастает. Однако растения батата боятся холода и неморозоустойчивы. Поэтому создание новых сортов и гибридов, обладающих устойчивостью к пониженным температурам является актуальной проблемой. Клеточная биотехнология направлена на решение данной проблемы с применением методов клонального микроразмножения, клеточной селекции, соматической гибридизации и др. Для быстрого размножения и получения высококачественного посадочного материала применяют методы биотехнологии, в частности, клональное микроразмножение. Однако в этой технологии существуют трудности, связанные с плохой адаптацией микроклонов к условиям ex vitro. Этот факт вносит дополнительное требование к подбору оптимальных режимов укоренения in vitro и адаптации ex vitro микроклонов.Материал и методика. Целью работы было изучение влияния условий культивирования на укоренение in vitro и адаптации ex vitro микроклонов I. batatas (L.). Объектом исследования были микрочеренки батата, размноженные in vitro. Микрочеренки I. batatas культивировали in vitro на питательной среде, содержащей минеральные соли по прописи Мурасига и Скуга, а также различные ауксины. Изучали влияние красного (R) и дальнего красного (FR) света на укоренение микропобегов in vitro и адаптацию микроклонов ex vitro.Результаты. Экспериментально установлено, что выращивание микрочеренков на питательной среде, содержащей ИМК в концентрации 0,5-1 мг/л и в условиях освещения светодиодными лампами красного и дальнего красного света в равном соотношении, приводит к получению микроклонов с хорошо развитой корневой системой и надземной биомассой. Применение аэропонной установки на последнем этапе клонального микроразмножения позволяет получать высококачественный посадочный материал, способный хорошо переносить адаптацию к условиям открытого грунта

Transplantation technologies for treatment of carbohydrate metabolism disorders

The review includes results of retrospective and prospective clinical studies (foreign and national) and guidelines on the use of transplantation technologies for treatment of type 1 diabetes and pancreatogenic diabetes in chronic pancreatitis and pancreatic conditions. Modern data on prevalence of diabetes and modern insulin delivery methods are presented. Results of transplantation of pancreas and islets of Langerhans in primary insulin-dependent conditions are considered. Analysis of the technology for isolation and autotransplantation of islets after pancreatectomy in chronic pancreatitis and benign tumor diseases are given

Monitoring membrane viscosity in differentiating stem cells using BODIPY-based molecular rotors and FLIM

Membrane fluidity plays an important role in many cell functions such as cell adhesion, and migration. In stem cell lines membrane fluidity may play a role in differentiation. Here we report the use of viscosity-sensitive fluorophores based on a BODIPY core, termed “molecular rotors”, in combination with Fluorescence Lifetime Imaging Microscopy, for monitoring of plasma membrane viscosity changes in mesenchymal stem cells (MSCs) during osteogenic and chondrogenic differentiation. In order to correlate the viscosity values with membrane lipid composition, the detailed analysis of the corresponding membrane lipid composition of differentiated cells was performed by time-of-flight secondary ion mass spectrometry. Our results directly demonstrate for the first time that differentiation of MSCs results in distinct membrane viscosities, that reflect the change in lipidome of the cells following differentiation

Label-free sorting of iPS cells during neuronal differentiation using FLIM and multiphoton fluorescence microscopy

The changes in cell metabolism can affect the epigenome-modifying enzymes activity during iPSCs differentiation and thus control the functional potential of the final cell. Therefore, for therapeutic applications, the restoration of a fully functional mitochondrial network specific for the cell types derived from iPSCs will be required to support the energy and other mitochondrial factors. Recently, FLIM method allows to study the metabolic changes that accompanying cell differentiation noninvasively and without additional labels. In this study, we investigated the metabolic changes in iPSCs during neural differentiation using two-photon fluorescence microscopy and FLIM. Cellular metabolism was examined by monitoring the optical redox ratio (FAD/NAD(P)H), the fluorescence lifetime contributions of the free and bound forms of NADH and NADPH. Given that neural differentiation is also accompanied by synthetic proceßes and oxidative streß, this proceß was included in the scope of this work. We demonstrated an increased contribution of protein-bound NADH and NADPH in neuron aßociated with metabolic switch to oxidative phosphorylation and the biosynthetic proceßes or oxidative streß, respectively. We also found that the optical redox ratio FAD/NAD(P)H decreased during neural differentiation, and this was likely to be explained by the intensive lipid membrane synthesis or ROS generating and the enhanced NADPH production aßociated with them. The biochemical analysis was carried out to verify the metabolic status of iPSCs and their neural derivatives. Based on the data on glucose consumption, lactate and ATP amount we registered the trend to the metabolic pathways redistribution towards the oxidative phosphorylation in neuron. © COPYRIGHT SPIE. Downloading of the abstract is permitted for personal use only

Probing metabolic alteration of differentiating induced pluripotent stem cells using label-free FLIM

The differentiation of endothelial cells from human iPSC has incontestable advantages in diseases research and therapeutic applications. However, the safe use of iPSC derivatives in regenerative medicine requires an enhanced understanding and control of factors that optimize in vitro reprogramming and differentiation protocols. Shifts in cellular metabolism associated with intracellular pH changes affect the enzymes that control epigenetic configuration, which impact chromatin reorganization and gene expression changes during reprogramming and differentiation. FLIM-based metabolic imaging of NADH and FAD is a powerful tool for measuring mitochondrial metabolic state and widely used diagnostic method for identification of neoplastic diseases, skin diseases, ocular pathologies and stem cells differentiation. Therefore, in this study, we used the potential of FLIM-based metabolic imaging and fluorescence microscopy of NADH and FAD to study the metabolic changes during iPSC differentiation in endothelial cells. The evaluation of the intracellular pH was carried out with the fluorescent pH-sensor SypHer-2 and fluorescence microscopy to obtain complete information about metabolic status of iPSC and their endothelial derivatives. Based on the FAD/NAD(P)H optical redox ratios increase and the contributions rise of the NAD(P)H fluorescence lifetime in iPSC during endothelial differentiation, we demonstrated an contribution increase of OXPHOS to cellular metabolism. Based on the shift toward more acidic intracellular pH in endothelial cell derived from iPSCs we verified their oxidative state. © COPYRIGHT SPIE. Downloading of the abstract is permitted for personal use only