"Citologiâ" publishes articles on all major problems of cell biology: morphology, physiology, immunology, biochemistry, molecular biology, biophysics. The journal accepts previously unpublished original articles on research of both animal and plant cells, reviews, discussion articles, reports on new research methods, reviews of books published in the current year, chronicles. Messages about scientific meetings (congresses, conferences, symposiums, etc.) for the "Chronicle" section of the journal are accepted only if they are submitted no later than 2 months from the date of the meeting.


The journal "Citologiâ" is abstracted/indexed in: eLIBRARY , Scopus , RSCI in Web of Science , Ulrich's Periodicals Directory .

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Vol 65, No 3 (2023)

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New in Centromere Genomics: Lessons from the First T2T Human Genome Assembly
Uralsky L.I., Alexandrov I.A., Ryabov F.D., Lapidus A.L., Rogaev E.I.

With the recent development of long-read sequencing technologies, it is now possible for the first time to read a complete gapless sequence of the human genome. The result was the first T2T (telomere-to-telomere) genomic assembly, published by an international consortium of scientists in 2022. The most significant contribution of the new assembly were the centromeric regions consisting of highly repetitive satellite DNA. In this review, we will briefly list the major achievements of the T2T consortium related to centromeres and take a closer look at the unexpected findings of cytogenetic magnitude that analysis of first assembled human centromeres has brought, such as the “split” centromeres of chromosomes 3 and 4, mega-inversion in the active centromere array of chromosome 1, haplotypic epialleles in the centromere of X chromosome and the macro-repeats found in several centromeres.

Citologiâ. 2023;65(3):217-231
pages 217-231 views
Ubiquitin–Proteasome System in Cell Pluripotency and Differentiation
Podenkova U.I., Zubarev I.V., Tomilin A.N., Tsimokha A.S.

Pluripotent stem cells (PSCs), represented primarily by embryonic stem cells and induced pluripotent stem cells (iPSCs), have a unique ability to self-renew and differentiate into all types of somatic cells. Dissecting molecular mechanisms controlling these properties is important for an efficient and safe introduction of PSCs into clinics. Growing evidence indicates that the proteostasis plays a central role in PSCs fate decisions. This review focuses on the role of the ubiquitin-proteasome system, a key member of the proteostasis network, in the regulation of pluripotency and differentiation of PSCs.

Citologiâ. 2023;65(3):232-245
pages 232-245 views
The Role of the Integrated Response of Tumor Cells to Stress, Autophagy, and Chaperones in the Origin of Recurrent Resistant Tumors
Zubova S.G., Gnedina O.O.

Chemotherapy and radiotherapy are a colossal stress factor for tumor cells. In response to therapy, the entire evolutionarily fixed response of cells to stress is activated. This happens at all levels of cell organization, namely at the protein level and the DNA level. This response involves the cell proteostasis system, DNA repair systems, tumor suppressor genes, and many other cell systems. We will consider the role of the main systems of proteostasis in these processes, namely, macroautophagy and chaperones, which are part of the integrated response of the cell to stress. As a result of the cell’s response to stress, the tumor cell becomes even less differentiated, activating the genes and intracellular systems necessary for survival. Cells that have responded to stress in this way have a more aggressive phenotype that is significantly more resistant to therapy. Under the influence of stress, the cell evolutionarily simplifies, which gives it additional chances for survival. On the one hand, autophagy contributes to a decrease in tumor cell differentiation and its plasticity, and on the other hand, it maintains a certain stability, being responsible for the integrity of the genome and freeing the cell from damaged organelles and defective proteins. Both autophagy and chaperones contribute to the acquisition of multidrug resistance by the tumor, which further complicates therapy. Understanding these processes makes it possible to develop new therapeutic approaches, taking into account the multistage nature of carcinogenesis.

Citologiâ. 2023;65(3):246-258
pages 246-258 views
Microglia Cell Line SIM-A9 Features – New Data
Shaposhnikova D.A., Moskaleva E.Y., Semochkina Y.P., Vysotskaya O.V., Komova O.V., Nasonova E.A., Koshlan I.V.

SIM-A9 is a line of spontaneously immortalized mouse microglia cells obtained from newborn C57BL/6 mice’s cerebrum. The aim of this work is to characterize SIM-A9 line by the ratio of cells with the resting and activated microglia phenotype, to analyze the expression of stem/progenitor cell markers CD133 and nestin, growth factors receptors CSF-1R and EGFR, and the karyotype of this line. The light microscopy, immunocytochemistry, flow cytometry and RT/PCR were used to analyze the morphology, phenotype, and gene expression levels of pro-inflammatory cytokines, and the mFISH method was used to analyze the karyotype. It was shown for the first time that SIM-A9 cells express a high level of TSPO protein, CD68, CD11b and CD45 markers on the surface membrane of cells, which corresponds to the phenotype of activated microglia. Despite this, the cells of this line respond with additional activation to LPS stimulation, which leads to an increase in the pro-inflammatory cytokine genes IL-1β, TNFα, IL-6 expression and a high level of active oxygen and nitrogen metabolites formation. It was shown that SIM-A9 cells express stem and progenitor cells markers, CD133+ and nestin, which allows us to consider the cells of this line as early poorly differentiated progenitor cells, despite their phenotype corresponding to activated microglia. It was also found that SIM-A9 cells express receptors of two growth factors CSF-1 and EGF, CSF-1R and EGFR, which indicates the possibility of SIM-A9 cells proliferation stimulation by two alternative mechanisms under the action of the corresponding factors. SIM-A9 cells have a hypotetraploid karyotype with a large number of structural and quantitative chromosome anomalies.

Citologiâ. 2023;65(3):259-272
pages 259-272 views
Impact of the Stimulation and Inhibition of NAD Biosynthesis on the Maintenance of Pluripotency in Mouse Embryonic Stem Cells
Antipova M.V., Kulikova V.A., Solovjeva L.V., Kropotov A.V., Svetlova M.P., Yakimov A.P., Nerinovski K.B., Bakhmet E.I., Nikiforov A.A.

Nicotinamide adenine dinucleotide (NAD+) plays a key role in cellular metabolism and signaling. In recent years, evidence has accumulated that NAD+-dependent processes are involved in the regulation of pluripotency and differentiation of mammalian embryonic stem cells. The major means to maintain NAD+ levels in mammalian cells is through its biosynthesis from various forms of vitamin B3. In this study, we examined how stimulation and inhibition of NAD+ biosynthesis affect the maintenance of the pluripotency of mouse embryonic stem cells E14 Tg2a (E14 cells). The pluripotency status of E14 cells was assessed by immunocytochemical and immunoblotting analysis using antibodies to the pluripotency factor Oct4, as well as by staining for alkaline phosphatase. Using NMR spectroscopy, we have found that the concentration of NAD+ in pluripotent E14 cells cultured in the presence of LIF is about 4 nmol/mg, and it remains unchanged after induction of differentiation with retinoic acid. We have also demonstrated that pharmacological stimulation of NAD+ biosynthesis by nicotinamide riboside increases the level of intracellular NAD+ by 20%, but it does not affect the maintenance of pluripotency in E14 cells. Moreover, under conditions of critical depletion of NAD+ pool by Nampt inhibition with FK866 E14 cells maintained pluripotency, though the expression level of Oct4 was decreased.

Citologiâ. 2023;65(3):273-282
pages 273-282 views
Titanium Dioxide Nanoparticles Inhibits Entosis in the Human Breast Adenocarcinoma Cell Line
Kisurina-Evgenieva O.P., Savitskaya M.A., Smeshnova D.S., Onishchenko G.E.

TiO2 is widely used in industry and cosmetics and medicines production. In recent years, to achieve tumor-specific delivery of anticancer agents, TiO2 nanoparticles have been used in chemo/photodynamic therapy, which may cause local increase of the TiO2 concentration in tumors. The TiO2 nanoparticles can affect various processes in tumors. One of such process is entosis. During entosis one tumor cell invades another tumor cell. The aim of this work was to study the effect of TiO2 nanoparticles (anatase <25 nm and rutil/anatase <75 nm; 1, 10 and 100 µg/mL, 72 hours) on the entosis in the human breast adenocarcinoma cell line (MCF7). Cultivation of cells in the presence of nanoparticles lead to a slowdown in proliferation and reduced in the entosis number. These effects were dose-dependent. Elemental analysis (analytical electron microscopy) showed presence TiO2 nanoparticles in the cell vacuoles, in the cytosol and in the extracellular space. TiO2 nanoparticles (10 µg/mL) significantly disrupted adhesive junctions in entotic cells and in cell culture in general (immunocytochemistry staining). The anatase nanoparticles induced p53 translocation into the nucleus. Thus, the obtained data showed that the TiO2 nanoparticles inhibited entosis in MCF-7 cells by means of disrupting the adhesive junction formation and preventing cell invasion. However, failure of adhesive contacts can facilitate tumor metastasis.

Citologiâ. 2023;65(3):283-294
pages 283-294 views
CRISPR/Cas9 Induced Duplications, Deletions and Inversions in Mouse Zygotes Lead to Karyotype Instability
Minina J.M., Soroka A.B., Karamysheva T.V., Serdyukova N.A., Serov O.L.

CRISPR/Cas9 technology has been widely used for targeted modification of the mammalian genomes. We have analyzed the karyotype of 18 mouse fibroblast cell lines with Cntn6 gene rearrangements introduced by CRISPR/Cas9. We have produced cell lines with 2374 kb Cntn6 gene duplications, 1137 kb deletions and inversions of similar size. In addition, we have performed cytogenetic analysis for five control mouse embryonic fibroblasts with the intact Cntn6 gene alleles. The cell lines heterozygous for Cntn6 gene inversion and homozygous and heterozygous for Cntn6 gene duplication had a high level of polyploidy (20–46%), as well as chromosome 6 monosomy (1–9%) and trisomy (1–8%). No trisomy was detected in the four cell lines with the deletion and duplication of the Cntn6 gene in the compound, and the proportion of polyploid cells was minimal (1.5–5.7%). Thus, we have shown the karyotype destabilization in the cell lines that have undergone genome editing using CRISPR/Cas9 system.

Citologiâ. 2023;65(3):295-302
pages 295-302 views
Visualization of Single Cells in the State of SOS Response Using Expansion Microscopy
Rumyantseva N.A., Golofeeva D.M., Vishnyakov I.E., Vedyaykin A.D.

Expansion microscopy (ExM) is a sample preparation method that allows to achieve improved visualization of structures due to the physical expansion of the sample. This method is used in combination with traditional light microscopy and allows, without the use of complex technical devices typical for super-resolution microscopy, to achieve visualization of biological structures with higher resolution. Unlike the methods of super-resolution microscopy, expansion microscopy does not make it possible to overcome the diffraction limit; however, the observed effect can be considered equivalent to an increase in the spatial resolution. The relative simplicity of the method and the undemanding nature of the microscope used have made expansion microscopy a fairly popular method to visualize various biological structures last time. This paper describes the use of expansion microscopy to visualize DNA and structures formed by the FtsZ protein in Escherichia coli cells during the SOS response. The results of the work confirm the previously obtained data that the FtsZ protein in cells in the state of the SOS response is unevenly distributed. The protocol used in this work for visualization of E. coli cells preliminarily fixed on the glass surface using the expansion microscopy method can be used in the future to study the internal structures of other cells, both bacterial and eukaryotic.

Citologiâ. 2023;65(3):303-310
pages 303-310 views

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