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  • Title: ANALYSIS OF THE INFLUENCE OF PROLONGED IRRADIATION ON HEMATOPOIETIC PROGENITOR CELLS IN GEL DIFFUSION CHAMBERS USING MATHEMATICAL MODELLING.
    Author: Bilko DI, Boiko RV, Russu IZ, Dyagil IS, Bilko NM.
    Journal: Probl Radiac Med Radiobiol; 2022 Dec; 27():203-215. PubMed ID: 36582090.
    Abstract:
    OBJECTIVE: determining of the functional activity of mice bone marrow hematopoietic progenitor cells, cultivated in gel diffusion chambers, on the stages of hematopoiesis recovery after their prolonged irradiation in the lethal dose in a comparative aspect with the method of colony forming in spleen using mathematical model. MATERIALS AND METHODS: The method of cell cultivation in gel diffusion chambers, cytological methods, mathematical modeling, and statistical methods of research were used. Bone marrow samples extracted from the femur of mice irradiated with a total dose of 8 Gy with a power 0.0028 Gy/min were cultivated in diffusion chambers with semi solid agar in the abdominal cavity of CBA recipient mice. RESULTS: Comparative analysis of the colonyforming efficiency of progenitor cells (CFU) was carried out during cultivation in gel diffusion chambers in the process of hematopoiesis recovery for 30 days, as well as in the spleen of lethally irradiated animals, in accordance with the mathematical model. Analysis of colony forming kinetics in gel diffusion chambers after prolonged exposure to ionizing radiation indicated the biphasic nature of hematopoiesis recovery. Thus, in the first few days after the irradiation a drop in the number of CFU is observed compared to the control, which continues until the 9th day. Subsequently there is a sharp increase in the number of CFU in cell culture, which continues until the complete recovery of hematopoiesis. The obtained data, recalculated per mouse femur, correspond to the results of colony forming in the spleen of irradiated animals, described by K. S. Chertkov and taken as a basis while developing our mathematical model, as well as to its parameters, which describe the process of hematopoiesis recovery. CONCLUSIONS: Conformity of the indices obtained during the cultivation using the method of gel diffusion chambers of mice bone marrow prolongedly irradiated at a total dose of 8 Gy with a power 0.0028 Gy/min, to the results of colony forming in spleen of lethally irradiated mice, which were the basis for mathematical model development, is the evidence of the feasibility of using a mathematical model to assess the process of hematopoiesis recovery by progenitor cells of different maturation levels, and the experimental approach of CFU growing in gel diffusion chambers can be considered as an additional method of researching the hematopoiesis recovery along with the spleen colony method. Meta: vyznachennia funktsional'noï aktyvnosti gemopoetychnykh klitynpoperednykiv kistkovogo mozku mysheĭ, kul'tyvovanykh u gelevykh dyfuziĭnykh kamerakh, na etapakh vidnovlennia gemopoezu pislia ïkh prolongovanogo oprominennia u letal'niĭ dozi v porivnial'nomu aspekti z metodom koloniieutvorennia u selezintsi z vykorystanniam matematychnoï modeli.Materialy i metody. Bulo vykorystano metod kul'tyvuvannia klityn u gelevykh dyfuziĭnykh kamerakh, tsytologichni metody, matematychne modeliuvannia, statystychni metody doslidzhen'. Zrazky kistkovogo mozku, vyluchenogo zi stegnovoï kistky mysheĭ i oprominenogo u sumarniĭ dozi 8 Gr z potuzhnistiu 0,0028 Gr/khv, kul'tyvuvaly u dyfuziĭnykh kamerakh iz napivridkym agarom u cherevniĭ porozhnyni mysheĭretsypiientiv liniï CBA. Rezul'taty. Provedeno porivnial'nyĭ analiz efektyvnosti koloniieutvorennia klitynpoperednykiv (KUO) pry kul'tyvuvanni u gelevykh dyfuziĭnykh kamerakh u protsesi vidnovlennia gemopoezu protiagom 30 dib, a takozh u selezintsi letal'no oprominenykh tvaryn, u vidpovidnosti do matematychnoï modeli. Analiz kinetyky koloniieutvorennia u gelevykh dyfuziĭnykh kamerakh pislia prolongovanoï diï ionizuiuchogo oprominennia svidchyv pro dvofaznyĭ kharakter vidnovlennia gemopoezu. Tak, u pershi kil'ka dib pislia oprominennia sposterigaiet'sia padinnia kil'kosti KUO porivniano z kontrolem, iake prodovzhuiet'sia do 9ï doby. Dali vidbuvaiet'sia rizke zrostannia kil'kosti KUO v kul'turi klityn, iake prodovzhuiet'sia do povnogo vidnovlennia gemopoezu. Otrymani dani, u pererakhunku na stegnovu kistku myshi, vidpovidaiut' rezul'tatam koloniieutvorennia u selezintsi oprominenykh tvaryn, opysanym K. S. Chertkovym (1973, 1977) ta vziatym za osnovu pry pobudovi namy matematychnoï modeli, a takozh ïï parametram, iaki opysuiut' protses vidnovlennia gemopoezu. Vysnovky. Vidpovidnist' pokaznykiv, otrymanykh pry kul'tyvuvanni metodom gelevykh dyfuziĭnykh kamer kistkovogo mozku mysheĭ, oprominenykh prolongovano u sumarniĭ dozi 8 Gr z potuzhnistiu 0,0028 Gr/khv, rezul'tatam koloniieutvorennia u selezintsi letal'no oprominenykh mysheĭ, shcho buly pokladeni v osnovu pobudovy matematychnoï modeli, svidchyt' pro dotsil'nist' vykorystannia matematychnoï modeli dlia otsinky protsesu vidnovlennia gemopoezu klitynamypoperednykamy riznykh rivniv dozrivannia, a eksperymental'nyĭ pidkhid vyroshchuvannia KUO u gelevykh dyfuziĭnykh kamerakh mozhe rozgliadatysia iak dodatkovyĭ sposib doslidzhennia vidnovlennia gemopoezu narivni z metodom selezinkovykh koloniĭ.
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