ПРОЯВЛЕНИЯ ГЕТЕРОГЕННОСТИ И ПОЛИМОРФИЗМА КЛЕТОК КАРЦИНОМЫ МОЛОЧНОЙ ЖЕЛЕЗЫ ПРИ КУЛЬТИВИРОВАНИИ

Abstract

The aim of the research was assessment of morphological characteristics of cells of two populations, isolated during the development of the method of cultivating a sample of breast carcinoma in order to create a personalized experimental model. Materials and methods. For our work, we used the surgical material of breast carcinoma. After preparation, tumor tissue was incubated in a mixed solution of enzymes, after which the sample was filtered and resuspended in a complete nutrient medium containing antibiotics-antimycotics. The cell suspension was placed in a seed vial with a nutrient medium and incubated in a CO2 incubator until a monolayer was formed. The culture medium was changed every 3 days. Subculture of cell lines was performed when the culture reached 80–90% confluent once every 6–7 days. Control over the state of the culture was carried out using an Eclipse TS100 inverted microscope, Nikon. To assess morphological parameters, cells were stained according to Romanovsky. The linear dimensions of the nuclei and the cells themselves were calculated using an MOV-1-15x ocular micrometer and a Micros MC50 light microscope. Micrographs of the preparations were made on a Leica DM5000 B microscope at 400 magnification. Statistical processing of the results was carried out using the MS Excel and STATISTICA 6 programs; the Mann — Whitney test was used to assess the significance of differences. Results. During the cultivation of breast carcinoma cells during three passages in the culture, two types of cells were identified — adhesive and non-adhesive, in which, with an increase in the cultivation time, the indicators of the relative cell area, the relative area of the nuclei and the nuclear-cytoplasmic ratio significantly increased. In addition, in these populations, the manifestation of polymorphism was noted with an increase in the cultivation time, which manifests itself in an increase in the proportion of larger, irregularly shaped cells at each subsequent passage. Conclusion. As a result of the study, it was revealed that the culture of breast carcinoma cells is characterized by the appearance in the culture of two populations of cells — adhesive and non-adhesive with high proliferative activity, which is confirmed by significant changes in cell sizes, nuclei, nuclear-cytoplasmic ratio. In addition, cell polymorphism is observed in both populations, which is accompanied by an increase in the proportion of larger irregularly shaped cells in culture with an increase in the cultivation time and the number of passages.