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Minimum Essential Medium (minimum + essential_medium)
Selected AbstractsEffect of temperature and storage media on human periodontal ligament fibroblast viabilityDENTAL TRAUMATOLOGY, Issue 3 2010Beatriz Dulcineia Mendes Souza The purpose of this study was to compare the effectiveness of several storage media to preserve cultured periodontal ligament fibroblasts (PDLF) under different temperatures. The media tested were: sterile Hank's balanced salt solution (sHBSS), non-sterile HBSS (nHBSS), skimmed milk, Save-A-Tooth®, Minimum Essential Medium (MEM) and water (negative control). MEM at 37°C was used as positive control. PDLF were obtained from explants of extracted healthy human teeth. Plates containing confluent PDLF were soaked in the various media for 3, 6, 24, 48 and 72 h at 37°C and 20°C. After incubation, viability of the cells was determined using the tetrazolium salt-based colorimetric (MTT) assay and the Trypan Blue exclusion test after 6, 24, 48 and 72 h of incubation at 20°C. The results were analyzed statistically using Kruskal,Wallis, Scheffé and Mann,Whitney (, = 5%) tests. Results from the MTT assay at 37°C and 20°C showed that skimmed milk was the best storage medium for up to 24 and 48 h, respectively, followed by nHBSS and sHBSS. Results from the Trypan Blue exclusion test showed that the best storage media were milk, sHBSS and nHBSS, with no statistical differences, for any time period. The Save-A-Tooth® had a detrimental effect on cells after 24 h. The influence of temperature on the effectiveness of the storage media tested showed at 20°C a decreasing order of efficacy as follows: milk > sHBSS and nHBSS > MEM > Save-A-Tooth® > water while at 37°C it was: MEM > nHBSS > milk > sHBSS > Save-A-Tooth® > water. In conclusion, incubation temperature altered the effectiveness of the storage media and skimmed milk at 20°C was better than HBSS in maintaining PDLF viability. [source] Overexpression of RGPR-p117 enhances regucalcin gene promoter activity in cloned normal rat kidney proximal tubular epithelial cells: Involvement of TTGGC motifJOURNAL OF CELLULAR BIOCHEMISTRY, Issue 2 2006Natsumi Sawada Abstract A novel protein RGPR-p117 was discovered as regucalcin gene promoter region-related protein that binds to the TTGGC motif using a yeast one-hybrid system. RGPR-p117 is localized in the nucleus of kidney cells, and overexpression of RGPR-p117 can modulate regucalcin protein and its mRNA expression in the cloned normal rat kidney proximal tubular epithelial NRK52E cells. This study was undertaken to determine whether overexpression of RGPR-p117 enhances the regucalcin promoter activity using the ,710/+18 LUC construct (wild-type) or ,710/+18 LUC construct (mutant) with deletion of ,523/,435 including TTGGC motif. NRK52E cells (wild-type) or stable HA-RGPR-p117/phCMV2-transfected cells (transfectant) were cultured in Dulbecco's minimum essential medium (DMEM) containing 5% bovine serum (BS). Wild-type cells or transfectants were transfected with the ,710/+18 LUC construct vector or the ,710/+18 LUC construct with deletion of ,523/,435. Wild-type cells or transfectants with subconfluency were cultured for 48 h in a DMEM medium containing either vehicle, BS (5%), or parathyroid hormone (1,34) (PTH; 10,7 M). Luciferase activity in wild-type cells was significantly increased with culture of BS or PTH. This increase was significantly blocked in the presence of various protein kinase inhibitors (staurosporine and PD 98059). Luciferase activity in transfectants was significantly increased as compared with that of wild-type cells in the absence of BS or PTH. The increase in luciferase activity in transfectants was completely decreased in mutant with deletion of ,523/,435 sequence of regucalcin promoter. This was also seen using the ,710/+18 LUC construct with deletion of ,523/,503 sequence containing TTGGC motif. The increase in luciferase activity in transfectants was not significantly enhanced with culture of BS (5%), PTH (10,7 M), Bay K 8644 (10,6 M), phorbol 12-myristate 13-acetate (PMA; 10,6 M), or N6, 2,-dibutyryl cyclic adenosine 3,, 5,-monophosphate (DcAMP; 10,4 M). The increase in luciferase activity in transfectants was completely inhibited with culture of dibucaine (10,6 M), staurosporine (10,9 M), PD 98059 (10,8 M), wortmannin (10,8 M), genistein (10,6 M), vanadate (10,6 M), or okadaic acid (10,6 M) which are inhibitors of various kinases and protein phosphatases. This study demonstrates that RGPR-p117 can enhance the regucalcin promoter activity which is related to the NF-1 consensus sequences including TTGGC motif, and that its enhancing effect is partly mediated through phosphorylation and dephosphorylation in NRK52E cells. J. Cell. Biochem. 99: 589,597, 2006. © 2006 Wiley-Liss, Inc. [source] Establishment and characterization of two new cell lines derived from flounder, Paralichthys olivaceus (Temminck & Schlegel)JOURNAL OF FISH DISEASES, Issue 11-12 2003M S Kang Abstract Two new cell cultures from flounder, Paralichthys olivaceus (Temminck & Schlegel), flounder fin (FFN) cells from fin tissue and flounder spleen (FSP) cells from spleen tissue, were established and characterized. The cells multiplied well in Eagle's minimum essential medium, supplemented with 10% foetal bovine serum, and have been subcultured more than 100 times, becoming continuous cell lines. Modal diploid chromosome number of FFN and FSP cells was 64 and 62, respectively. Polymerase chain reaction products were obtained from FFN and FSP cells with primer sets of microsatellite markers of flounder. Optimal growth temperature was 20 °C and consisted of epithelioid cells. FFN and FSP cells showed cytopathic effects after inoculation of infectious pancreatic necrosis virus, marine birnavirus, chum salmon virus, infectious haematopoietic necrosis virus, spring viraemia of carp virus and hirame rhabdovirus. Thus these new cell lines may be useful for studying a wide range of fish viruses. [source] Anti-influenza virus activity of crude extract of Ribes nigrum L.PHYTOTHERAPY RESEARCH, Issue 2 2003Yoko M. Knox Abstract This experiment was designed to detect the antiviral activities of crude fruit extracts of wild Ribes nigrum L. (Kurokarin extract) against influenza virus types A and B. Kurokarin extract was prepared as follows: fruits of Ribes nigrum L. were heated at 50,°C in a heating tank, and then ground under anaerobic conditions. The extracts were centrifuged, and the supernatant fluid was filtered and sterilized by infrared rays. The crude extract was diluted with Eagle's minimum essential medium (MEM) and the solution was adjusted to a pH 7.2 with 0.1 N or 1 N NaOH. Proven anti-influenza virus effects of the extracts were shown. The concentration of extract required to inhibit the plaque formation of both IVA and IVB by 50% (IC50) was 3.2,,g/mL. Both IVA and IVB were directly inactivated up to 99% by 10,,g/mL of the extract at pH 2.8, and 95% to 98% by this dose at pH 7.2. The growth of IVA in cells treated with 10 and 100,,g/mL of the extract for 6,h after infection was completely suppressed. Virus titres in culture fluids of the cells treated with 100,,g/mL of Kurokarin extract for 1,h at 8 to 9,h after infection, were completely suppressed, indicating that the extract inhibited the virus release from the infected cells. Copyright © 2003 John Wiley & Sons, Ltd. [source] | ||||||||||