Aim. To study the influence of fetal bovine serum and serum of adult rats on behavior of newborn rat isolated neural cells during their cultivation in vitro. Methods. The isolation of neural cells from neonatal rat brain. The determination of the dynamics of cellular monolayer formation. Immunocytochemical staining of cells for b-tubulin III, nestin and vimentin. Results. It has been determined that the addition of serum of adult rats to the cultivation medium creates more favorable conditions for survival, attachment and spread of differentiated, and proliferation of the stem/progenitor neural cells of newborn rats during cultivation in vitro compared with the fetal bovine serum. Conclusions. Using the serum of adult rats is preferable for the cultivation of isolated neural cells of newborn rats compared with the fetal bovine serum.
According to WHO, 6.7 million people die from stroke every year. The search for new neuroprotective substances remains an urgent task.The purpose of this study was to investigate the neuroprotective activity of factors of placental origin.Materials and methods. Neuroprotective activity of media conditioned with cryopreserved placenta derived mesenchymal stem cells (MSCs), organotypic culture of placenta and placental extract was studied on in vitro models of glutamate excitotoxicity in rats` neural cells. Neural cells were cultured with placental factors without glutamate treatment, before and after glutamate treatment. Neural cells` metabolic activity was assessed by MTT test.Results. Placental factors increase the MTT test indexes, prevent the toxic effect of glutamate on neural cells and promote their recovery. The thermolability of factors of placental origin and the effectiveness of various placental preparations are shown.Conclusions. Conditional media of placenta derived MSCs, organotypic culture of the placenta and human placental extract have neuroprotective effect on rats` brain cells in vitro.
This review discusses the characteristics of three-dimensional cell culture systems on and without carriers (scaffolds). Scaffolds are used to simulate the extracellular matrix, as well as to reproduce the natural physical and structural microenvironment of cells, similar to living tissue. The review examines the types of scaffolds (hard and gel-like, natural and artificial, degradable and non-degradable), their characteristics, advantages and disadvantages, features of cell distribution in them. The use of decellularized and devitalized organs and tissues as scaffolds is discussed. The review also considers matrix-free cultivation of cells in the composition of three-dimensional multicellular structures -spheroids. The structure and biology of spheroids is discussed. The features of spheroid formation under static (self-assembly) and dynamic (under the influence of external forces) cultivation conditions are considered. The role of spheroid size for cell survival is discussed.
Background. The human placenta is a promising source of biomaterial for regenerative medicine. This is primarily due to the availability of a sufficient amount of material, low immunogenicity, a large number of stem cells, and high proliferative cell potential. The effectiveness of stem cells, their lysates, and conditioned media in various pathological conditions has been proven in many studies. Most studies are experimental, or are in different phases of clinical trials. At the same time, in Ukraine there is a wealth of experience of clinical using the cryopreserved placenta medico-immunobiological preparations, which can be the basis for further application of placental material. Objective. We aim to perform a critical analysis of data on the effectiveness and prospects of application of the cryopreserved placenta preparations (cells, tissues, membranes, extract and cord blood serum) in clinical practice. Methods. The results of clinical application of 2,579 medical immunobiological Platex and Cryocell placenta preparations were analyzed. An attention was paid to the effectiveness, course of concomitant, comorbid pathology in patients, as well as the presence of negative responses and complications. The number of the preparations used and the one of the patients according to the nosological forms were counted. Results. The experience of application of medical immunobiological preparations of placental origin in obstetric-gynecological, therapeutic, neurological and endocrinological pathologies is analyzed in the work. The obtained data are compared with the research results in the corresponding model experiments of in vitro and in vivo systems. The positive effect of placental preparations on the course of miscarriage, climacteric syndrome, infertility, diabetes mellitus, coronary heart disease, multiple sclerosis, amyotrophic lateral sclerosis, trophic ulcers has been determined. Contraindications for placenta preparations are some malignancies and an infectious disease process without proper pathogen elimination. Conclusions. Cryopreserved placenta preparations are effective when used in obstetric, gynecological, neurological, endocrinological and therapeutic practice. Their effect is primarily observed in diseases, which are accompanied by autoimmune reactions, hormonal disorders, dysplastic or degenerative processes. The restricted application of placenta preparations is infectious disease process without proper pathogen elimination.
High efficacy of placental cells application necessitates their investigation. Preclinical studies require an improvement of the methods for obtaining, standardizing and storage of placental cells of experimental animals. Cells were isolated from rats and mice placentas by means of different enzymatic methods and the one of explants. Cells were cryopreserved with DMSO in DMEM using two-stage freezing. The number, morphological, cultural, metabolic features of cells were studied after isolation and storage. The maximum number of viable cells from the placentas of mice and rats was found to be obtained using the explant method or trypsin with ETDA. Cell cultures from mice and rats placentas after the third passage had stable morphofunctional characteristics. The viability of warmed rat placental cells according to dye exclusion was (92.3 ± 1.6)%, according to the adhesive test this was (81.3 ± 5.8)%. For mice placental cells, these values were (86.7 ± 3.7)% and (79.2 ± 8.1)%, correspondingly. The research results enabled the determining of effective biotechniques for obtaining the cryopreserved placental cells of rats and mice to perform preclinical studies of their biological effect in models of allo- and autotransplantations.
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