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Introduction. Despite significant advances in the creation of stable cell lines, the focus of research has recently shifted toward the creation of primary cell cultures derived directly from patient tumor samples, which include both tumor cells and microenvironmental cells.The aim of the study was to compare the morphological characteristics of the cells of a breast carcinoma sample when cultured over three passages.Materials and methods. Material for the study was obtained during surgical intervention in a patient diagnosed with breast carcinoma. Slices were prepared from the tumor sample according to the standard histological protocol and stained with monoclonal antibodies to estrogen, progesterone, Ki-67, Her2/neu receptors. Cell nuclei were stained with hematoxylin. Immunohistochemical reaction was performed in DAKO autostainer (Denmark). Part of the material was placed in Hanks' solution with 5% antibiotic antimycotics and delivered to the Cell Culture Laboratory, where after performing the standard protocol for obtaining cell culture, tumor cells were diluted in Mammocult nutrient medium and placed in culture vials. For morphological evaluation, cells were stained by Pappenheim. For immunocytochemical analysis in determining the belonging of cells to epithelial cells using anti-Pan Keratin Primary Antibody antibody. The number of cells was counted in an automatic TC20 counter, and culture growth was monitored using an Eclipse TS100 microscope, Nikon (Japan).Results and Discussion. On the basis of immunohistochemical study, the tumor sample was classified as Luminal-A subtype. During the study several groups of cells were isolated and cytologically evaluated. The results of immunocytochemical analysis of the cultured cells confirm that the tumor cells retained their epithelial phenotype during culturing. In spite of the manifestation of cell polymorphism in BML cell culture, during three passages the cultured tumor cells retained their epithelial nature and showed a tendency to form a monolayer.Conclusion. A detailed study of cytomorphology and immunocytological characteristics of cultured cells of different immunohistochemical PBMC subtypes will help to evaluate the main regularities of tumor cell vital functions in vitro and allow a more differentiated approach to the creation of personalized cell cultures in order to develop a targeted chemotherapeutic effect on tumors of specific patients. |
