Comparison of phenotype change in cells of fat tissue, bone marrow and pancreatic gland of cats with passages
Journal Title: Бюлетень "Ветеринарна біотехнологія" - Year 2017, Vol 31, Issue
Abstract
Introduction. Considering that one of the important parts of the pathogenesis of diabetes mellitus is the death of beta-cells and due to the disadvantages of modern methods of treating diabetes mellitus 1 with the use of insulin therapy and pancreatic gland (PG) tissue transplantation, there is a stimulation of the development and improvement of cell therapy methods.Therefore, our study is a comparison of changes in phenotypic cell characteristics in order to create an experimental system for evaluating and selecting the optimal source of cellular material for further treatment of diabetes mellitus.The goal of the work Compare phenotypic changes in fat tissue, bone marrow and pancreatic gland of cats from the first to the fifth passage.Materials and methods. In the experiment fat tissue, bone marrow and pancreatic gland were used to produce cell cultures. The material was received in parallel during scheduled surgical operations. The control of phenotype changes was performed by detecting CD markers (CD10, CD38, CD34, CD45, CD48, CD54, CD56, CD66e, CD96, CD227, CD326, CD pan keratin). Theresults were analyzed by the number of cells with expression (green glow of cells) and evaluated using the classic H-Score method.Results of research and discussion. Studies have shown that the primary cell cultures of these tissues are morphologically heterogeneous, they include: a small number of polygonal cells diffusely located among the fibroblast-like cells, which constituted the main percentage of cell culture. It should be noted that the ratio of polygonal cells to fibroblast-like cells in the studied cultures was different. At a later stage of cultivation, we observed increase in the percentage of fibroblast-like cells, which led to the homogenization of the cell composition of cultures with passages. Investigation of the immunophenotype of cell cultures of bone marrow, fat tissue and pancreatic gland also showed differences that did not disappear during the cultivation.Conclusion and prospects for further research. The results of our studies have shown that the primary culture of adhesive cells of all studied cultures consists of different types of cells, due to the diversity of the output pool.Within passages, an increase in the percentage of fibroblast-like cells in the studied cell cultures was observed.During passage, the CD profile of cells in the studied cultures varied, with certain regularities. Based on the range of used CD markers specific for non-specialized (low- differentiated) cells, the lowest expression level was observed in the cells cultures of pancreatic gland, insignificantly higher level in cells cultures of fat tissue (compared with cells cultures of bone marrow),which can indicate a higher level of cell culture universality received from bone marrow. This hypothesis is confirmed by the deceleration of a monolayer formation in cultures of pancreatic gland and fat tissue, while in the cells cultures of bone marrow, the time of achievement of the confluence remained stable.A different phenotype cells culture of bone marrow, fat tissue and pancreatic gland will allow using them in cellular therapy in the future and it is expected they influence differently under the same conditions.
Authors and Affiliations
Anatoly Mazurkevich, Vitaliy Kovpak, O. Kovpak, Natalia Hudz
Keywords
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- EP ID EP266485
- DOI 10.31073/vet_biotech31-11
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