03. Науково-дослідний інститут експериментальної та клінічної медицини ХНМУ

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    Isolation and Cryopreservation of Placental Cells: Search for Optimal Biotechniques in Experimental and Regenerative Medicine
    (2021) Prokopiuk, O.; Shevchenko, M.; Prokopiuk, V.; Musatova, I.; Safonov, R.; Prokopiuk, O.
    Abstract: A high effi cacy 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 diff erent 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. 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 eff ective biotechniques for obtaining the cryopreserved placental cells of rats and mice to perform preclinical studies of their biological eff ect in models of allo- and autotransplantations.
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    The study of phospholipid bilayer of cell membranes in leukocytes incubated with high concentrations of the food additive E407a
    (2021) Tkachenko, Anton; Onishchenko, Anatolii; Roshal, Alexander; Posokhov, Yevgen
    Aim: The aim of the study was to assess the impact of semi-refined carrageenan (E407a) on the hydrophobic region of phospholipid bilayer in cell membranes of leukocytes exposed to the solution with high concentrations of this food additive. Material and methods: Fluorescent probe (2-phenylphenanthro[ 9,10-d]-1,3-oxazole) was used to investigate the influence of E407a on the state of lipid bilayer in leukocytes extracted from rats and treated with a 5% solution of the food additive E407a during 4 hours. Results: The shapes of the probe fluorescence spectra did not differ in leukocytes of rats treated with the 5% solution of semi-refined carrageenan and white blood cells of control samples. Such findings suggest that exposure to the E407a solution causes no changes in the proton-donor ability of the media in the lipid membranes of leukocytes in the region where the probe locates. Conclusion: Exposure of white blood cell suspensions to the semirefined carrageenan solution does not affect the membrane hydration of the hydrophobic region of leukocyte phospholipid bilayer.
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    Effects of semi-refined carrageenan (food additive E407a) oncell membranes of leukocytes assessed in vivo and in vitro
    (2021) Tkachenko, Anton; Onishchenko, Anatolii; Roshal, Alexander; Nakonechna, Oksana; Chumachenko, Tetyana; Posokhov, Yevgen
    Aim: To assess the impact of semi-refined carrageenan (E407a) on hydrophobic regions of phosphololipid bilayer in cell membranes of leukocytes collected from rats orally administered this food additive and white blood cells incubated with E407a. Methods: Fluorescent probes (ortho-hydroxy derivatives of 2,5-diaryl-1,3-oxazole) were used to estimate the state of lipid bilayer in leukocytes obtained from rats orally exposed to the food additive E407a and in white blood cells incubated with E407a. Results: No noticeable changes in the physico-chemical properties were observed in the lipid membranes of leukocytes in the region where the probes locate in response to oral intake of semi-refined carrageenan. Incubation of leukocytes with E407a solutions resulted in a decrease in polarity and proton-donor ability of leukocytes in the area of carbonyl groups of phospholipids and in the area of hydrocarbon chains of phospholipids near the polar region of the bilayer. Conclusion: Membrane fluidity abnormalities found in cells exposed to E407a are similar to those observed in patients with IBD suggesting that contribution of carrageenan to IBD development may be partially explained by leukocyte membrane modifications.