Cytological evaluation of a single cell culture of Luminal A subtype breast carcinoma cells

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.

Effects of Oxytetracycline and Gentamicin Therapeutic Doses on Hematological, Biochemical and Hematopoietic Parameters in Cyprinus carpio Juveniles

Hematological, biochemical and hematopoietic effects of therapeutic doses of two antibiotics, oxytetracycline (OTC) and gentamicin (GEN), in clinically healthy common carp juveniles were studied. The fish were divided into four groups: controls 1 and 2 (untreated or injected with 0.6% NaCl solution), and two groups treated with antibiotics (orally with 75 mg/kg OTC four times every two days or injected with a single dose (4 mg/kg) of GEN dissolved in 0.6% NaCl). Blood and head kidneys were sampled from all fish 3 days post-treatments for hematological, biochemical and hematopoietic tissue analyses. No major alterations in the values of hematological and serum biochemical parameters occurred following administration of OTC or GEN. Glucose concentrations were significantly lower in both groups of fish subjected to injections (Control 2 and GEN), while the oxidative metabolic activity of phagocytes increased in the antibiotic-treated groups (significantly in OTC). More alterations were observed in hematopoietic tissue. Immunocytochemical analysis revealed that G caused a significant increase in the rate of cell proliferation (PCNA-positive cells) and an increase in the frequency of apoptotic cells (caspase-positive). The frequency of lymphoid lineage decreased, which was related to a decrease in the abundance of mature lymphocytes in GEN-treated fish. Percentages of neutrophilic lineage were significantly elevated in OTC and GEN groups compared to controls. The obtained results showed no considerable hematotoxicity or hepatotoxicity of therapeutic doses of OTC and GEN to carp.

Rhodopsin and melanopsin coexist in mammalian sperm cells and activate different signaling pathways for thermotaxis

Recently, various opsin types, known to be involved in vision, were demonstrated to be present in human and mouse sperm cells and to be involved there in thermosensing for thermotaxis. In vision, each opsin type is restricted to specific cells. The situation in this respect in sperm cells is not known. It is also not known whether or not both signaling pathways, found to function in sperm thermotaxis, are each activated by specific opsins, as in vision. Here we addressed these questions. Choosing rhodopsin and melanopsin as test cases and employing immunocytochemical analysis with antibodies against these opsins, we found that the majority of sperm cells were stained by both antibodies, indicating that most of the cells contained both opsins. By employing mutant mouse sperm cells that do not express melanopsin combined with specific signaling inhibitors, we furthermore demonstrated that rhodopsin and melanopsin each activates a different pathway. Thus, in mammalian sperm thermotaxis, as in vision, rhodopsin and melanopsin each triggers a different signaling pathway but, unlike in vision, both opsin types coexist in the same sperm cells.