RESUMEN
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.
Asunto(s)
Hematopoyesis , Células Madre Hematopoyéticas , Ratones , Animales , Ensayo de Unidades Formadoras de Colonias , Ratones Endogámicos CBA , Células Madre Hematopoyéticas/efectos de la radiación , Hematopoyesis/efectos de la radiación , Radiación IonizanteRESUMEN
OBJECTIVE: development of the humanized system for cells cultivation outside the human organism (human-mouse)and investigation of the influence of ionizing radiation in increasing doses on the colony-forming ability ofhematopoietic progenitor cells. MATERIALS AND METHODS: Bone marrow samples of individuals without blood system diseases were cultivated in geldiffusion chambers with semi-solid agar in the abdominal cavity of CBA mice exposed to ionizing radiation action.Cell aggregates, which were obtained in the culture of diffusion chambers in vivo, were counted and colony-formingefficiency of bone marrow cells was determined. RESULTS: We revealed the stimulation of colony forming under the action of ionizing radiation in increasing doseson the animals-recipients of the chambers, which indirectly indicates the synthesis of colony-stimulating factor inthe mice organism and its permeation into the diffusion chambers with human bone marrow cells. The effect of cyto-statics action on the mice organism was investigated, which in experimentally selected dose cause stimulation ofcolony forming in cell cultures, both 24 hours and 2 hours after administration. CONCLUSIONS: The ability of hematopoietic progenitor cells of bone marrow to form colonies and clusters was eval-uated during the cultivation in semi-solid agar in gel diffusion chambers in vivo, as well as the association with thenumber of explanted cells in the appropriate range was established, which indicates the clonal nature of cell aggre-gates growth in culture. It was shown that the treatment of animals the day prior to experiment with administra-tion of cytostatics is comparable to the action of ionizing radiation and can be used to study hematopoiesis in«human-mouse¼ system.
Asunto(s)
Proliferación Celular/efectos de la radiación , Hematopoyesis/efectos de la radiación , Células Madre Hematopoyéticas/efectos de la radiación , Dosis de Radiación , Radiación Ionizante , Animales , Modelos Animales de Enfermedad , Humanos , Ratones , Ratones Endogámicos CBARESUMEN
OBJECTIVE: To perform comparative analysis of the characteristics of population functioning process of mice bonemarrow colony-forming units after their prolonged irradiation in lethal and non-lethal doses with equal dose rateintensity with the aid of mathematical model. MATERIALS AND METHODS: Assigned task is solved by means of mathematical model of alterations in the number ofbone marrow colony-forming units after continuous irradiation, described in previous works, with the use of experimental results of K. S. Chertkov works (1972, 1973). Mathematical model is developed basing on the hematopoiesisscheme introduced by I. L. Chertkov (1984, 1991). RESULTS AND CONCLUSIONS: By applying original mathematical model, new scheme of hematopoiesis [6], with theuse of experimental results of γ-irradiation influence in the doses of 4 Gy and 8 Gy with the dose rate intensity of0.0028 Gy/min on the number of mice bone marrow colony-forming units, as well as experimental data concerningthe processes of their number recovery, obtained from literature references, we determined the parameters, whichcharacterize hematopoietic system reaction on the different stages of recovery processes of mice bone marrowcolony-forming units after the termination of ionizing radiation action; comparative analysis of obtained resultswas performed.
Asunto(s)
Células de la Médula Ósea/efectos de la radiación , Médula Ósea/efectos de la radiación , Hematopoyesis/efectos de la radiación , Células Madre Hematopoyéticas/efectos de la radiación , Modelos Teóricos , Traumatismos por Radiación/patología , Animales , Médula Ósea/patología , Células de la Médula Ósea/patología , Quimera , Ensayo de Unidades Formadoras de Colonias , Relación Dosis-Respuesta en la Radiación , Femenino , Rayos gamma/efectos adversos , Células Madre Hematopoyéticas/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos CBA , Traumatismos por Radiación/mortalidad , Análisis de Supervivencia , Irradiación Corporal TotalRESUMEN
OBJECTIVE: to determine the quantitative characteristics of population functioning of mice bone marrow colony-forming units during seven days of acute fractionated irradiation. MATERIALS AND METHODS: Assigned task is solved by means of described in works R. V. Boiko et al. (2015, 2016) math-ematical model of alterations in the number of bone marrow colony-forming units using the experimental results ofwork W.Chu-Tse, L. G.Lajtha (1975). Mathematical model is developed basing on the new hematopoiesis scheme,which was introduced by I. Chertkov(1984, 1991). RESULTS: By applying original mathematical model of hematopoiesis scheme using results concerning the change innumber of bone marrow colony-forming units of mice femur we determined quantitative characteristics of theirfunctioning during seven days of fractionated irradiation under daily acute γ-radiation in the dose of 0.7 Gy. CONCLUSIONS: Mathematical model is introduced, which describes changes in the relative number of colony-formingunits in mice bone marrow in the process of their acute fractionated irradiation.