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1

Abnormal cytokine production by bone marrow stromal cells of multiple myeloma patients in response to RPMI8226 myeloma cells

100%
Zdzisinska B., Bojarska-Junak A., Dmoszynska A., Kandefer-Szerszen M.
Archivum Immunologiae et Therapiae Experimentalis
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2008
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vol. 56
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issue 3
207-221
EN
Introduction Recent studies indicate that bone marrow stromal cells (BMSCs) derived from patients with multiple myeloma (MM) differ from those of healthy donors in their expression of extracellular matrix compounds and in cytokine production. It is not known whether these abnormalities are primary or are acquired by BMSCs on contact with MM cells. Materials and Methods: Interleukin (IL)-6, IL-11, IL-10, and tumor necrosis factor (TNF)-alpha production by CD166+ mesenchymal BMSCs and the CD38+/CD138+ RPMI8226 myeloma cell line cultivated in vitro in monocultures or co-cultivated under cell-to-cell contact or non-contact conditions in the presence of a tissue culture insert were measured. Intracellular cytokines were measured by flow cytometry analysis as the percentage of cytokine-producing cells or by mean fluorescence intensity as the level of cytokine expression in cells. Additionally, ELISA was used to measure IL-6, soluble IL-6 receptor (sIL-6R), IL-11, IL-10, TNF- alpha, B-cell-activating factor of the TNF family (BAFF), hepatocyte growth factor (HGF), and osteopontin (OPN) production in the supernatants of the cultures and co-cultures. Results A higher ability of the BMSCs of MM patients than in controls was detected to produce IL-6, IL-10, TNF- alpha, OPN, and especially HGF and BAFF in response to the RPMI8226 cells. Moreover, the BMSCs of the MM patients significantly enhanced the production of sIL-6R by the RPMI8226 cells. Discussion Cytokines over-expressed by BMSCs of MM patients can function as growth factors for myeloma cells (IL-6, IL-10, HGF), migration stimulatory factors for tumor plasma cells (TNF-alpha, HGF), adhesion stimulatory factors (HGF, BAFF and OPN), stimulators of osteoclastogenesis (IL-6, TNF-alpha), and angiogenic factors (TNF-alpha). The results of this experiment strongly suggest that the BMSCs from MM patients differed in spontaneous and myeloma cell-induced production of cytokines, especially of HGF and BAFF, and these abnormalities were both primary and acquired by the BMSCs on contact with the MM cells. This in turn suggests the presence of an undefined, autocrine stimulation pathway resulting in a prolonged production of cytokines even in long-term cultures in vitro and in vivo. These abnormalities might provide optimal conditions for the proliferation and differentiation of residual tumor cells or their precursors in the affected bone marrow.
2

Matrix metalloproteinase and cytokine production by bone marrow adherent cells from multiple myeloma patients

100%
Zdzisinska B., Walter-Croneck A., Dmoszynska A., Kandefer-Szerszen M.
Archivum Immunologiae et Therapiae Experimentalis
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2006
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vol. 54
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issue 4
289-296
EN
Introduction: Cultures of bone marrow stromal cells derived from the bone marrow of multiple myeloma (MM) patients were shown to exhibit several abnormalities compared with control cultures from healthy subjects. The aim of the study was to examine whether cultures of bone marrow adherent cells, at low passage level, exhibit differences in matrix metalloproteinases (MMPs) and cytokine production compared with cultures from normal donors. Materials and Methods: MMP production was evaluated by gel zymography and by ELISA in supernatants of serum-free cultures of bone marrow adherent cells derived from 20 MM patients and 23 healthy controls. Spontaneous and lipopolysaccharide (LPS)- or Newcastle disease virus (NDV)-induced cytokine release was assessed in the supernatants of the cultures by the ELISA method. Results: Both cultures produced MMP-1, -2, -3, and -9 under serum-free conditions; however, the levels of MMP-1 and MMP-2 were significantly higher in cultures derived from MM patients, while MMP-3 was significantly higher in control cultures. The level of MMP-9 was comparable in the cultures derived from MM patients and controls. All cultures produced interleukin (IL)-10 and IL-11 spontaneously, but after LPS or NDV induction the levels of IL-10, IL-11, interferon , and tumor necrosis factor , were significantly higher in the cultures derived from MM patients than in control cultures. Conclusions: The results indicate that both the abnormalities in MMP production and the overproduction of cytokines (in the presence of LPS or virus, which mimic inflammatory conditions) may be involved in bone destruction and tumor spread in multiple myeloma.
3

Gelatin sponge Spongostan as a carrier for animal and human adherent cell culture

88%
Paduch R., Rzeski W., Zdzisinska B., Kaczor J., Kandefer-Szerszen M.
Biotechnologia
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2003
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issue 1
223-233
EN
A porous gelatin sponge Spongostan (Ferrosan Denmark) as a carrier for the cultivation of several adherent cell lines and strains was tested. This material provide considerably greater surface available for cells than conventional bottles. Moreover, it allows to maintain high cell viability (over 87%) during culture time. When cells were cultured on Spongostan discs (wet vol. 0.11 cm3), no phenotypic changes in cell adherence and proliferation dynamics were observed. However, when cells adhered to Spongostan were frozen down and kept for 5 months in liquid nitrogen, they lost about 65% of their viability. Generally, Spongostan can be considered as a universal carrier for three-dimensional culture of many cell types, but it cannot be used for cells? freezing.
4
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Cell death and neuronal arborization upon quercetin treatment in rat neurons

88%
Jakubowicz-Gil J., Rzeski W., Zdzisinska B., Dobrowolski P., Gawron A.
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EN
Quercetin, one of the major flavonoids, exhibits many beneficial effects on human organism as antihistamine, antioxidant, anti-inflammatory, anticancer and antiviral drug. It is recommended as suplement of healthy diet but still the knowledge of its beneficial effect on normal cells is not satisfactory. We decided to examine the effect of flavonoid on neurons morphology and their susceptibility to cell death. Fractal analysis of rat neurons revealed that 24 hours long incubation with quercetin diminished neuronal arborisation in cortical neurons. Neurons also appeared to be very sensitive to cell death after flavonoid treatment in concentration dependent manner. Over 50% of cells died after incubation with 15 ?g/ml of flavonoid while 1 ?g/ ml of quercetin induced cell death only in 5%. Staining with Hoechst 33342 and propidium ioidide revealed the two types of cell death: apoptosis and necrosis. The number of apoptotic cells was comparable with necrotic ones. These results suggest toxic effect of quercetin on neurons what should be taken into consideration in further studies on using quercetin as therapeutic agent.
5
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Different sensitivity of neurons and neuroblastoma cells to quercetin treatment

76%
Jakubowicz-Gil J., Rzeski W., Zdzisinska B., Piersiak T., Weiksza K., Glowniak K., Gawron A.
Acta Neurobiologiae Experimentalis
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2008
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vol. 68
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issue 4
463-476
EN
determined the sensitivity of neurons and neuroblastoma cells on apoptosis and necrosis induction upon quercetin treatment. No expression of Hsp72 was observed in neurons, which were more sensitive to cell death upon quercetin treatment than neuroblastoma cells, where Hsp72 expression was observed. Reduction of Hsp72 gene expression in neuroblastoma cells by antisense oligonucleotides made them more sensitive to pro-apoptotic action of quercetin. Moreover, the flavonoid decreased Hsp27, procaspase-3, MRP and PKB expression in neuroblastoma cells and in neurons. Nuclear localization of mainly cytoplasmic Hsp27 was observed in neuroblastoma cells after treatment with high quercetin concentrations, while in neurons, the protein was present in nuclei both in control and quercetin treated cells. Our results suggest that quercetin induce apoptosis more effectively in cells with low level of Hsp 72 expression. Higher sensitivity of neurons for cell death after treatment with high quercetin concentrations in comparison to neuroblastoma cell line should also be taken into consideration in further studies on using studied flavonoid as therapeutic agent.
6

Propagation of Derzsy disease virus in goose embryo fibroblasts. An application of the Celligen Plus bioreactor

64%
Rzeski W., Paduch R., Kozdrun W., KaczorJ., Czekaj H., Samorek-Salamonowicz E., Kandefer-Szerszen M., Zdzisinska B., Dawidowicz A.
Biotechnologia
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2003
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issue 2
261-267
EN
Derzsy desease is a fatal hepatitis of young geese under the age of 5 weeks. The etiological agent is a goose parvovirus (GPV) which is widespread in all major goose farming countries in Europe and Asia. At present, vaccination is the only method preventing the loss of animals. In vitro GPV replicates only in goose embryo fibroblasts (GEF) and this way the virus is propagated in order to obtain the material for diagnostic tests and vaccine production. In this paper, we report the application of bioreactor Celligen Plus for GEF culture and Derzsy disease virus propagation. The method is based on immobilization and culture of cells on porous glass carriers in bioreactor. Comparing to the traditional cell culture techniques, the described method was over 12-fold more efficient and allowed to obtain good quality virus pool for Derzsy disease vaccine preparation.
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