Home About us Contact
|
Home About us Contact | ||
|
|
||
Eagle's Medium (eagle + medium)
Selected AbstractsC-peptide constricts pancreatic islet arterioles in diabetic, but not normoglycaemic miceDIABETES/METABOLISM: RESEARCH AND REVIEWS, Issue 2 2008Lina Nordquist Abstract Background Pancreatic islet blood flow is regulated separately from that of the exocrine pancreas, and a consistent finding during impaired glucose tolerance is an increased blood perfusion. The aim of the present study was to investigate whether C-peptide affects pancreatic islet arterioles in normal and diabetic mice. Materials and Methods Control and diabetic C57-Bl mice were studied after 2 weeks of alloxan-induced diabetes. Islet arterioles were dissected and microperfused with Dulbecco's modified Eagle medium (DMEM) solution. The effect of luminal application of mouse C-peptide was investigated. Results C-peptide reduced the diameter of islet arterioles from diabetic mice (,10 ± 4%, P < 0.05) compared to base-line values, whilst arterioles from normoglycaemic animals did not respond to C-peptide (P = 0.2). Conclusion These findings suggest a role for C-peptide in the regulation of islet blood flow, especially during conditions with impaired glucose tolerance. Copyright © 2007 John Wiley & Sons, Ltd. [source] Localization of substance P-induced upregulated interleukin-8 expression in human dental pulp explantsINTERNATIONAL ENDODONTIC JOURNAL, Issue 2 2008G. T.-J. Abstract Aim, To localize ex vivo expression of interleukin-8 (IL-8) induced by substance P (SP) in human dental pulps. Methodology, Intact caries-free, freshly extracted third molars (n = 20) were collected from patients (15,25 years old). The teeth were split and pulpal tissue was obtained and stored in Dulbecco's modified Eagle medium. Human dental pulp tissue explants were stimulated with SP. Expression of IL-8 in pulp explants was detected and localized by immunohistochemistry. Results, Moderated IL-8 immunoreactivities were detected mainly in the cell-rich zone in pulp tissues 12 h after tumour necrosis factor alpha (TNF-,) stimulation (positive controls), whereas only weak IL-8 expression was observed in tissues stimulated with SP at the same time interval. These data did not differ from those in negative controls. Increased IL-8 expression in pulp explants after 24 h of SP stimulation was noted compared with negative controls and located in fibroblast-like cells, blood vessel-associated cells and extracellular matrix in the central zone and cell-rich zone of pulp explants. Tissues stimulated with TNF-, for 24 h (positive controls) revealed weak IL-8 immunoreactivities with altered cell morphology. Conclusions, Substance P induces IL-8 expression and was located in fibroblast-like pulp cells, blood vessel-associated cells and extracellular matrix of human dental explants. These data support the hypothesis that neuropeptide (SP) coordinates the modulation of pulpal inflammation via up-regulating chemokine IL-8. [source] In Vitro Culture and Differentiation of Buffalo (Bubalus bubalis) SpermatogoniaREPRODUCTION IN DOMESTIC ANIMALS, Issue 2 2010B Xie Contents The objective of this study was to develop a culture system which could support buffalo spermatogonia differentiation into spermatids in vitro. Testes from 3- to 5-month-old buffaloes were decapsulated and seminiferous tubules were enzymatically dissociated to recover spermatogonia and sertoli cells. The cells were cultured in modified Dulbecco modified Eagle medium supplemented with different concentrations of foetal bovine serum, retinol, testosterone for 2 months at 37°C. Spermatogonia and sertoli cells were identified with an antibody against c-kit or GATA4, respectively. The viability of spermatogonia in the media supplemented with different concentrations of serum was all significantly higher (p < 0.05) compared with that in the medium without serum. A-paired or A-aligned spermatogonia and spermatogonial colonies (AP-positive) were observed after 7,10 days of culture and spermatid-like cells with a flagellum (6,8 ,m) appeared after 30 days of culture. For cultured conditions, retinol could not significantly promote the formation of spermatid-like cells (p > 0.05), whereas supplementation of testosterone could significantly promote (p < 0.05) the formation of spermatid-like cells after 41 days of culture. The expression of the spermatid-specific marker gene (PRM2) was identified after 30 days of culture by RT-PCR. Yet, the transition protein 1 (TP1, a haploid makers) was not detected. Meanwhile, spermatids developed in vitro were also confirmed by Raman spectroscopy. These results suggest that buffalo spermatogonia could differentiate into spermatids in vitro based on the analysis of their morphology, PRM2 expression and Raman spectroscopy. Yet, the normality of the spermatid-like cells was not supported by TP1 expression. [source] Organ preservation solutions attenuate accumulation and nuclear translocation of hypoxia-inducible factor-1, in the hepatoma cell line HepG2CELL BIOCHEMISTRY AND FUNCTION, Issue 8 2009Renate Paddenberg Abstract Hypoxia-inducible factor-1, (HIF-1,) is a key transcription factor orchestrating hypoxic and inflammatory reactions. Here, we determined the impact of organ preservation solutions (Celsior; histidine-tryptophan-ketoglutarate solution, HTK; University of Wisconsin solution; UW), oxygen supply, and temperature on HIF-1, accumulation, recorded by Western blotting and immunocytochemistry, in the human hepatoma cell line HepG2. Generation of reactive oxygen species (ROS), NO, and cell viability were concomitantly assessed. At 4°C, HIF-1, accumulation was not detectable. In normothermic (37°C) cell culture medium (Dulbecco's Modified Eagle's Medium, DMEM), HepG2 cells accumulated HIF-1, even in normoxia (21% O2) which was not observed in either of the preservation solutions. This correlated to high generation of NO, a normoxic stabilizer of HIF-1,, and L -arginine content (substrate for NO synthesis) in DMEM, and low NO production and absence of L -arginine in preservation solutions. In normothermic hypoxia up to 24,h, intracellular HIF-1, accumulated in all conditions, but less in preservation solutions compared to DMEM. The inhibitory effect on accumulation and nuclear translocation was most prominent for HTK, the only solution containing the activator of HIF-1, degradation, , -ketoglutarate. Addition of other intermediates of the tricarbon acid cycle,succinate, fumarate, malate,did not alter HIF-1, accumulation, although succinate exhibited a beneficial effect on cell viability in cold storage. In conclusion, preservation solutions attenuate accumulation and nuclear translocation of the transcription factor HIF-1,, and this property is seemingly related to their chemical composition (L -arginine, , -ketoglutarate). Thus, it appears feasible to design preservation solution specifically to modify HIF-1, accumulation and nuclear translocation. Copyright © 2009 John Wiley & Sons, Ltd. [source] Enhancement of In Vitro Hair Shaft Elongation in Follicles Stored in Buffers That Prevent Follicle Cell ApoptosisDERMATOLOGIC SURGERY, Issue 1 2004Walter Krugluger MD Background. Viability and survival of stored micrografts during hair follicle transplantation are important limitations of micrograft transplantation procedures. In this study, we investigated the effect of different storage solutions and inhibitors of apoptotic cell death (ACD) on hair follicle cell viability by measuring in vitro hair shaft elongation (HSE) for 5 days. Methods. Micrografts from informed patients undergoing routine micrograft transplantation were stored for 5 hours at room temperature in phosphate-buffered salt solution (PBS) or HEPES-buffered Dulbecco's modified Eagle's medium (DMEM), containing different concentrations of the ACD-inhibitors aminoguanidine (AMG), hormones (insulin, hydrocortisone), 14,15-epoxy-eicosatrienoic acid (14,15-EET), or combinations of these. Results. In vitro, HSE was significantly increased in micrografts stored in DMEM compared with PBS (2.3%±0.6% vs. 28.4%±3.9%, P<0.0001). DMEM supplemented with AMG (10 ,g/mL) or 14,15-EET (1 ng/mL) further increased in vitro HSE (33.9%±7.1%, p=0.01, and 32.8%±6.1%, P=0.02, respectively). Evaluation of ACD in stored micrografts, performed by determination of cytoplasmic histone-associated DNA fragments, confirmed the results found by HSE. ACD was detectable after a 36-hour culture in serum-containing medium and was higher in micrografts stored in PBS compared with micrografts stored in DMEM (A405nm/A492nm: 1.63±0.21 vs. 1.42±0.07, respectively; P<0.01). The addition of AMG further decreased serum-induced ACD in the micrografts (DMEM 1.42±0.07 vs. DMEM/AMG 0.90±0.11, P<0.0001). Conclusion. Our study demonstrated an important role of ACD in micrograft transplantation surgery. Preconditioning of micrografts with storage buffers containing inhibitors of ACD could prevent serum-induced ACD after transplantation and might increase the viability of micrografts and the clinical outcome in micrograft transplantation. [source] Cytotoxicity and genotoxicity of sodium percarbonate: a comparison with bleaching agents commonly used in discoloured pulpless teethINTERNATIONAL ENDODONTIC JOURNAL, Issue 2 2010M. R. Fernández Fernández MR, Carvalho RV, Ogliari FA, Beira FA, Etges A, Bueno M. Cytotoxicity and genotoxicity of sodium percarbonate: a comparison with bleaching agents commonly used in discoloured pulpless teeth. International Endodontic Journal, 43, 102,108, 2010. Abstract Aim, To evaluate the cytotoxicity and genotoxicity of sodium percarbonate (SPC) in comparison with bleaching agents used on discoloured pulpless teeth. Methodology, The cytotoxicity and genotoxicity of bleaching agents were evaluated both in their pure form as well as at concentrations commonly used in clinical practice. Hydrogen peroxide (HP), carbamide peroxide (CP), sodium perborate (SP) and SPC were diluted in Dulbecco's modified Eagle's medium (DMEM) in series. To evaluate the cytotoxicity, the survival of 3T3/NIH mouse fibroblasts was measured photometrically using an 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay after a 24 h-exposure period. Genotoxicity was indicated by micronuclei (MN) formation, and modification of the normal cell was analysed by light microscopy (400×). Statistical analysis was performed by one-way anova, followed by a multiple-comparison Tukey post hoc test (P < 0.05). Results, All groups exhibited a dose-dependent cytotoxicity. However, CP showed a similar cytotoxic effect when compared with DMEM-untreated control (UC) group. HP and SPC were significantly more cytotoxic than SP. The genotoxicity test showed that SPC and SP had an intermediate rate of MN frequency when compared with the UC group. The mean rate of MN frequency for HP was higher and statistically more significant than for the other groups tested. No difference was observed when CP and UC groups were compared. Conclusions, Sodium percarbonate showed cytotoxicity and genotoxicity similar to those of the other products tested. However, before SPC is used clinically, studies should be conducted to confirm its safety in vivo. [source] The effect of the ionic products of Bioglass® dissolution on human osteoblasts growth cycle in vitroJOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE, Issue 4 2007Jun-Ying Sun Abstract In this study, in order to observe the effect of Bioglass® and its ionic products on human esteoblasts growth cycle in vitro, the ionic products of Bioglass have been introduced to a cell culture medium by dissolving Bioglass particles in Dulbecco's modified Eagle's medium (DMEM) at 37 °C for 24 h; this was used as the experimental medium, while DMEM without Bioglass modification was used as the control medium. Human osteoblasts isolated from trabecular bone were treated by the two media and the timing of the osteoblast growth cycle was examined. Cell growth curves were derived after 7 days. Also, human osteoblasts were treated for 1,6 days by the two media, and the G1, S, G2 phase percentages of osteoblasts were recorded by flow cytometry every day, resulting in the cell proliferation activity index: SPF (S-phase fraction) and PI (proliferation index). The difference in cell growth was shown after the second day of culture (p < 0.01), and cell growth in the experimental groups was greater than in control groups. The SPF and PI of the experimental groups were also higher than the control groups in 2 days of culture (p < 0.05 and p < 0.01), which indicates that the growth cycle of the human osteoblasts in experimental medium is about 2 days. In conclusion, Bioglass can promote osteoblast proliferation, reducing the human osteoblast growth cycle to pass through G1 and S phase and then enter G2 phase quickly. Copyright © 2007 John Wiley & Sons, Ltd. [source] Ability of low-molecular-weight heparin to alleviate proteinuria by inhibiting respiratory syncytial virus infectionNEPHROLOGY, Issue 7 2008YANNAN GUO SUMMARY: Aim: Low-molecular-weight heparin (LMWH) is a negatively charged glycoprotein and has a very similar structure to that of cell surface heparin sulfate (HS). Thus, LMWH, an analog of HS, may inhibit positively charged respiratory syncytial virus (RSV) infection through cooperative electrostatic association. Methods: In this study, rats were respectively treated with 400 IU/kg LMWH before, during or after being inoculated with 6 × 106 plaque-forming unit (PFU) RSV. RSV and normal control groups were respectively inoculated by RSV and virus-free Dulbecco's modified Eagle's medium (DMEM). HeLa cells in vitro were pretreated with LMWH, elastase (ELA), heparinase (HpaIII) and protamine before being inoculated with 6 × 101 PFU RSV. RSV infectivity was determined by in situ hybridization and plaque assay. Results: After inoculation, the urinary protein excretion and serum parameters in LMWH-treated rats were significantly lower than those in the RSV group. No abnormalities of glomerular structure were observed in LMWH-treated groups whereas swelling and slight hypercellularity in minority glomeruli and foot process effacement were observed in the RSV group. RSV RNA of LMWH-treated rats had weaker expression than that of the RSV group. In vitro, RSV infection in RSV + LMWH, HpaIII + ELAI, protamine + ELAI, ELAI, HpaIII and protamine treatment cells were significantly lower than that of the RSV control, and that in RSV + LMWH was the least. There were no significant differences in RSV infection between ELAI + LMWH and RSV control. Conclusion: Our study confirmed that there is a correlation between RSV and proteinuria in rats. LMWH can alleviate proteinuria in rats through inhibiting RSV from binding with HS which plays an important role in the onset of RSV infection. [source] Effects of FSH and LH on Steroid Production by Buffalo (Bubalus bubalis) Granulosa Cells Cultured In Vitro Under Serum-Free ConditionsREPRODUCTION IN DOMESTIC ANIMALS, Issue 5 2010M Shanmugam Contents The objective of this study was to examine the effects of FSH and LH on oestradiol-17, and progesterone production by buffalo granulosa cells cultured under serum-free conditions. Granulosa cells (3 × 105) from small (,5 mm diameter) follicles were cultured for up to 4 days in 48-well plates coated with 3.3 ,g/cm2 fibronectin in Dulbecco's modified Eagle's medium (DMEM) : nutrient mixture F-12 Ham (1 : 1 ratio) supplemented with 10,7 m androstenedione, 5 ,g/ml human apo-transferrin and 0.1% bovine serum albumin, in the presence or absence of FSH or LH (0, 1, 2, 4, 8, 16, 32 or 64 ng/ml each). Basal oestradiol-17, production by granulosa cells from small follicles reduced (p < 0.01) from days 1 to 2 of culture and became undetectable by day 3 and basal progesterone production increased (p < 0.05) from day 1 through day 4 of the culture. Although there was no effect of FSH on day 1 of the culture, FSH at 2, 4, 8 and 16 ng/ml increased (p < 0.05) oestradiol-17, production by granulosa cells from small follicles on day 2. Progesterone secretion was increased (p < 0.05) by all doses of FSH on all days of culture. All doses of LH had no effect on oestradiol-17, or progesterone production by granulosa cells from small follicles on any day of the culture. The results of this study demonstrate a serum-free culture system for buffalo granulosa cells and stimulatory effect of FSH but not LH on steroid hormone production by buffalo granulosa cells under these conditions. [source] Primary cell cultures from anaplastic thyroid cancer obtained by fine-needle aspiration used for chemosensitivity testsCLINICAL ENDOCRINOLOGY, Issue 1 2008Alessandro Antonelli Summary Objective, Anaplastic thyroid cancer (ATC) is often inoperable and chemotherapy and radiotherapy are the main treatments. Until now, ,primary ATC cell cultures' (ANA) have been developed from surgical biopsies. We investigated the possibility of obtaining ANA from fine-needle aspiration (FNA-ANA) and testing their sensitivity to chemotherapeutic agents, which could enable treatments to be more effective and avoid unnecessary surgical procedures. Design and patients, The aim of this study was to obtain FNA-ANA from three ATC patients and to evaluate the chemosensitivity of FNA-ANA to chemotherapeutic agents. Measurements and results, FNA-ANA from ATC patients were cultured in RPMI 1640 and propagated in Dulbecco's modified Eagle's medium (DMEM). Chemosensitivity was evaluated by inhibiting the proliferation (analysing the number of viable cells by the cleavage of tetrazolium salts), by increasing the concentration of four different chemotherapeutic agents: bleomycin, cisplatin, gemcitabine and etoposide. The chemotherapeutic agents significantly inhibited (> 50%) FNA-ANA proliferation. Another ANA for each patient was obtained from a surgical biopsy specimen; the results for the chemosensitivity tests were similar to those obtained using FNA-ANA. Conclusions, Our study demonstrates the possibility of obtaining FNA-ANA, and opens the way to the use of FNA-ANA as a means of testing the chemosensitivity to different chemotherapeutic agents (and possibly the radiosensitivity) in each patient, avoiding unnecessary surgical procedures and the administration of inactive chemotherapeutics. [source] Tissue engineering of periosteal cell membranes in vitroCLINICAL ORAL IMPLANTS RESEARCH, Issue 8 2009Patrick H. Warnke Abstract: Objectives: The cultivation of bone is a major focus in tissue engineering and oral implantology. Without a periosteal layer, instant or rapid development of a substantial cortical layer is unlikely for engineered bone grafts. The aim of this study was to test the ability of four collagen membranes to support and promote the proliferation of human periosteal cells. Materials and methods: Human periosteum cells were cultured using an osteogenic medium consisting of Dulbecco's modified Eagle's medium supplemented with fetal calf serum, penicillin, streptomycin and ascorbic acid at 37°C with 5% CO2. Four collagen membranes served as scaffolds: Bio-Gide, Chondro-Gide, Tutodent and Ossix Plus. Cell vitality was assessed by fluorescin diacetate (FDA) and propidium iodide (PI) staining, biocompatibility with LDH and BrdU, MTT, WST tests and scanning electron microscopy (SEM). Results: After 24 h, all probes showed viable periosteal cells. All biocompatibility tests revealed that proliferation on all membranes after treatment with eluate from membranes after a 24-h immersion in a serum-free cell culture medium was similar to the controls. Periosteal cells formed layers covering the surfaces of all four membranes 7 days after seeding in SEM. Conclusion: It can be concluded from our data that the collagen membranes can be used as scaffolds for the cultivation of periosteum layers with a view to creating cortical bone using tissue-engineering methods. [source] | ||||||||||