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CELL CULTURES
Information Bulletin Issue 29, St. Petersburg, 2013 |
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THE PROBLEM OF GENOMIC STABILITY IN HUMAN EMBRYONIC STEM CELLS
The review cites a literature data about genetic stable and unstable continuous human embryonic stem sell (hESC) lines. The reasons of genetic instability appearance are viewed and the analyses of methods for studding a genetic stability are carried out. The examples of karyotype changes, some of which has an adaptation character, are contained in. The analysis of certain adaptive karyotype changes, which appear during long term cultivating and support self renewal of hESC from one side and malignancy from another, are carried out. Considering the perspective of utilization hESC lines in applied biomedical research, it is necessary to do the genetic stability control of hESC lines by cytogenetic and molecular methods of different sensitivity. COMPARATIVE CHARACTERIZATION OF STROMAL CELL CULTURES FROM HEMATOPOIETC ORGANS
OF RAT AT DIFFERENT ONTOGENETIC STAGES
Mesenchymal stromal cells (MSC) represent a crucial component of hematopoietic stroma. An analysis of their phenotypic and functional features at different developmental stages allows elucidating the process of establishing the hematopoietic system. The aim of this study was to compare morphology, immunophenotype and proliferative activity of MSC derived from fetal and adult rat hematopoietic organs in primary and passed cultures. Stromal cells were isolated from fetal liver at 16 and 20 days of development, spleen of 20-day-old fetuses and adult rats, femur of 20-day-old fetuses with developing bone marrow, and adult bone marrow. The cells of all examined organs formed discrete colonies when plated with a density of 1õ106 cells per milliliter. The only exceptions were the fetal femur cells that formed a confluent monolayer. Most of clonogenic cells from the liver of 16-day-old embryos, in contrast to the adult bone marrow, were sensitive to hydroxyurea, and therefore were in proliferative cycle. Primary cultures of fetal liver, spleen and femur as well as adult bone marrow contained cells that expressed CD73, CD90 and CD106 antigens characteristic for MSC. Expression of these antigens continued at the first passage. Passed cultures derived from all sources consisted of both fusiform cells and flat fibroblasts whose morphology was typical for senescent cells. In the culture of adult bone marrow and spleen numbers of flat cells increased at the third and second passages, respectively, but this did not occur in the cultures of fetal liver and spleen. The proportion of proliferating cells derived from hematopoietically active liver and spleen was higher compared with the cultures of these organs and bone marrow at the later developmental stages. Thus, the stromal cells of all studied sources had similar morphology and phenotype, but differed in proliferative activity. Key words: mesenchymal stromal cells, fetal liver, bone marrow, spleen, clonal growth, immunophenotype, proliferative activity COMPARATIVE STUDY OF CAPACITY FOR THE DIFFERENTIATION OF CULTIVATED NEURAL STEM
CELLS FROM THE HUMAN OLFACTORY BULB
Phenotypical peculiarities of neural cells from human olfactory bulb (OB) natal tissue have been researched on histological preparation and in dissociated cultures. Under the conditions of long-term cultivation in comparative aspect a differential potential of OB neural cells has been explored depending on the composition of cultivating environment. By cultivating in nutritious environment with a serum, the authors detected formation of multicellular spheroid units containing neural cells with positive reaction on vimentin that reflected their progenitor potential. By cultivating in nutritious environment without a serum the survival of most neural cells decreases considerably (in 9-10 times by the 30th day), here with in part of neural cells short primitive sprouts are detected as a demonstration of initial signs of spontaneous neural differentiation. By cultivating neural cells from OB in nutritious environment without a serum in the daytime with retinoic àñid the authors detected cytological signs of neural cells differentiation with terminal formation of neurocytes of multipolar shape with long dendritic sprouts creating network-like structures. The presence of retinoic àñid in cultivating environment also provides high survival of neural cells in comparison with other modifications of their cultivating that apparently reflects the protective properties of this preparation. Thus, in dissociated cultures of neural cells from human OB the authors detected their ability to neural differentiation, which severity depends on the composition of nutritious environment. Key words: neural stem cells, olfactory bulb, neural cells culturing, neural differentiation THE EFFECT OF THE PROLIFERATION INHIBITOR CYTOSINE-ARABINOSIDE ON THE CARDIOMYOCYTE
CONTRACTILE APPARATUS ORGANIZATION
Cardiomyocyte contractile apparatus rearrangement upon culturing thereof in the absence of the proliferation inhibitor cytosine-arabinoside is described, and cytosine-arabinoside is shown not to have an essential effect on the culturing cardiomyocyte contrañtile apparatus rearrangement pattern and not to be the reason thereof. Key words: cardiomyocytes, contractile apparatus, cytosine-arabinoside CYTOTOXIC ACTIVITY OF FRESHLY ISOLATED SPLENOCYTES OF MICE AND RATS AGAINST TUMOR
CELLS OF MONOLAYER CULTURES
Natural killer (NK) cells and immature dendritic cells (DC) are spontaneously cytotoxic immune effector cells (EC). To selectively destroy tumor cells NK cells utilize two main cytotoxicity pathways - perforin/granzyme-mediated secretory killing and TNF family mediated apoptotic killing, while DC only TNF-mediated apoptosis. In this study splenocytes of mice and rats were used as source of EC of innate immunite and monolayer cell lines of rat HTC and Zajdela, mouse MH-22a and BWTG3, and human HepG2 hepatomas and human A431 epidermoid carcinoma (Russian Cell Culture Collection, Institute of Cytology, Russian Academy of Sciences) - as target cells. It was shown, that mice splenocytes didn't induce the death of target cells of monolayer target cultures, whereas the splenocytes of rats demonstrated the distinct cytotoxic effect against hepatoma cells by using both secretory and apoptotic mechanisms of killing. Single intraperitoneal injected cyclophosphamide produced a immunomodulating effect on the activity of rat splenocytes - suppressor action on cells of Zajdela hepatoma and immunopotentiating action on the target cells of HTC, BWTG3, MH-22a and HepG2 hepatomas. Key words: apoptosis, cyclophosphamide, cytotoxic activity, hepatoma, monolayer cell cultures, natural killer cells, splenocytes, tumor cells INTERLEUKIN-6 - NEW MARKER OF RESISTANCE OF HUMAN NEUROEPITHELIAL TUMORS TO
CHEMOTHERAPEUTICAL DRUGS AND NERVE GROWTH FACTOR
The secretion of interleukin-6 (IL-6) of 83 neuroepithelial tumors cells (pilocytic, anaplastic astrocytomas, ependymomas, glio- and medulloblastomas) and their sensitivity to chemotherapy (carboplatin, methotrexate, cytarabine, cisplatin, etoposide) and nerve growth factor (NGF) ware studied in vitro. Application of the cytostatic drugs on the cell cultures of neoplasias strengthened (P <0,05) the loss of their cells in 92.0% of tests. Exposure of cultures with neurotrophin stimulated (P <0,05) of death in 80% of tumor types. Treatment of the cells of intracranial neoplasms with drugs reduced (P <0,05) in 56.0%, stimulated (P <0,05) in 4.0% and did not change in 40.0% of cases secretion of IL-6. Application of NGF reduced the (P <0,05) in the release of cytokines 100.0% of samples. Detected in 88.0 and 60% negative correlations (P <0,05) in the drug sensitivity (r = -0,815) and NGF (r = -0,596) cell neoplasias respectively, and secretion of IL-6. The use of IL-6 will allow clinicians to assess the effectiveness of the treatment and methods of its correction. Key words: IL-6, cell culture, neuroepithelial tumors, chemotherapeutical drugs, nerve growth factor, chemosensitivity DEGREE OF RESISTANCE TO LOW TEMPERATURES OF TUMOR CELLS IN COMPARISON WITH
BONE MARROW STEM CELLS
Current study focuses on cold tolerance of two types of tumor cells: human epidermoid carcinoma (line A 431) and freshly isolated tumor cells from mice (EHS sarcoma).
A comparative analysis of the results with the previous study about the resistance to low temperatures of normal bone marrow stem cells isolated from rats is relevant, considering the
similarity between tumor and stem cells, specifically their ability to self-renewal and active proliferation.
COMPARATIVE ANALYSIS OF EXTRACELLULAR MATRIX COMPONENTS SYNTHESYS BY HUMAN
FIBROBLASTS OF DIFFERENT ORIGIN DIRING CULTIVATING ON DIFFERENT SUBSTRATES
In given work the qualitative and quantitative assessment of extracellular matrix components synthesized by human dermal fibroblasts of different origin at cultivating on substrates with different adhesiveness was carried out. It was shown that fibroblasts isolated from normal and scar human tissue synthesize different amounts of type I collagen, fibronectin, chondroitin sulphate and decorin. In addition, it is possible that the rate of cells adhesion to substrate influences the process of extracellular matrix components synthesis. Key words: extracellular matrix proteins, perfluorodecalin, glycosaminoglycans, proteoglycans, fibroblasts COMPARE STUDY OF KARYOLOGIC CHARACTERISTICS VERO(B) CELL CULTURE RESTORE AFTER
THE LONG KRYOCONSERVATION AND ITS CULTIVATION IN DIFFERENT CULTURE MEDIA
It was showed, that after the long keeping (28 years) in liquid nitrogen the cell culture Vero(B) preserve stable karyology and morphology. Vero(Â) cell cultures were propagated during 10 passages in experimental culture medià, containing hydrolizate of soybean powder prepared using trypsin and bromeline enzymes, hydrolizate of rice powder prepared with trypsin, and in routine media (control). In that time observed change of the number of chromosomes. Experimental media supplied with 3% of fetal bovine serum showed high grow-enhancing properties and did not alter morphology of Vero (B) cell culture. Thus replace of routine mediums on more cheap ingredients wish 3% of fetal bovine serum essential didn't change biologic characteristics of cell culture Vero(B). Key words: cell cultures, culture media, keeping in nitrogen, karyology, morphology, prolypheration index |
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