Home About us Contact
|
Home About us Contact | ||
|
|
||
Eagle's Minimal Essential Medium (eagle + minimal_essential_medium)
Selected AbstractsNutrients Released by Gastric Epithelial Cells Enhance Helicobacter pylori GrowthHELICOBACTER, Issue 6 2004Karin Van Amsterdam ABSTRACT Background.,Helicobacter pylori survives and proliferates in the human gastric mucosa. In this niche, H. pylori adheres to the gastric epithelial cells near the tight junctions. In vitro, H. pylori proliferated well in tissue-culture medium near gastric epithelial cells. However, in the absence of epithelial cells, growth of H. pylori could only be established in tissue-culture medium when, prior to the experiment, it was preincubated near gastric epithelial cells. Therefore, we aimed to determine whether diffusion of nutrients derived from epithelial cells was required for H. pylori growth in Dulbecco's modified Eagle's minimal essential medium (DMEM) cell culture medium. Materials and Methods., Cell culture conditions essential for H. pylori growth in vitro were determined with gastric epithelial HM02 cells. Results., Deprivation of iron in cell-culture-conditioned DMEM resulted in a growth arrest of H. pylori. However, near gastric epithelial cells, growth of H. pylori was resistant to iron deprivation. Evidently, when residing close to epithelial cells, H. pylori was able to fulfil its iron requirements, even when the DMEM was deprived of iron. Nevertheless, supplementation with iron alone did not restore H. pylori growth in DMEM, hence other nutrients were deficient as well in the absence of epithelial cells. Growth of H. pylori in DMEM was restored when hypoxanthine, l -alanine and l -proline were added to the DMEM. Conclusions, Diffusion of (precursors of) these nutrients from the gastric epithelial cells is essential for H. pylori growth in vitro. We hypothesize that in vivo, H. pylori favors colonization near the tight junctions, to gain maximal access to the nutrient(s) released by gastric epithelial cells. [source] Evaluation of human nasal RPMI 2650 cells grown at an air,liquid interface as a model for nasal drug transport studiesJOURNAL OF PHARMACEUTICAL SCIENCES, Issue 3 2008Shuhua Bai Abstract This study tests the hypothesis that human nasal RPMI 2650 cells grown at an air,liquid interface is a feasible model for drug transport studies via the nasal route. RPMI 2650 cells were cultured in Eagle's minimal essential medium (MEM) at both air,liquid and liquid,liquid interfaces. For each culture regimen, monolayer integrity was tested by measuring the transepithelial resistance (TEER) as well as the transport of paracellular and transcellular markers across the monolayer. The expression of tight junction proteins,differentiation markers,in cells of the different monolayers was studied by western blot analysis and confocal microscopy. The highest TEER values (192,±,3 ,,·,cm2) were observed for RPMI 2650 cells seeded onto collagen-coated permeable polytetrafluoroethylene inserts and grown at an air,liquid interface for 10 days; a seeding density of 4,×,105/cm2 generated and maintained a cell monolayer with suitable barrier properties at days 9,12. Microscopic examination showed that RPMI 2650 cells grown on filter inserts formed a fully confluent monolayer. The apparent permeability coefficients of the paracellular marker, [14C] mannitol, and the transcellular marker, [3H] propranolol, were 5.07,±,0.01,×,10,6 cm/s and 16.1,±,0.1,×,10,6 cm/s, respectively. Western blot analysis indicated the presence of four tight junction proteins: ZO-1, occludin, claudin-1 and E-cadherin; and the quantities of ZO-1, occludin, and E-cadherin were significantly higher in cells grown at an air,liquid interface than in cells grown at a liquid,liquid interface. Confocal microscopic studies showed ZO-1, F-actin, occludin and claudin-1 proteins at cell-cell contacts and revealed significant differences in the distributions and densities of ZO-1 protein in cells grown at the two types of interface. The data indicate that RPMI 2650 cells grown at an air,liquid interface form polarized monolayers with the cells interconnected by tight junction proteins. This human nasal cell line model could provide a useful tool for in vitro screening of nasal drug candidates. © 2007 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 97:1165,1178, 2008 [source] Establishment and Characterization of a Normal Melanocyte Cell Line Derived from Pig SkinPIGMENT CELL & MELANOMA RESEARCH, Issue 4 2003Sophia Julé Several minipig strains develop spontaneous malignant melanoma. As a first step toward the analysis of genes involved in the tumoral progression of melanoma in these animal models, we developed culture conditions for pig melanocytes whereby melanocytes from normal epidermis can be isolated directly onto mitotically inactivated keratinocytes in Eagle's minimal essential medium supplemented with fetal calf serum, tetradecanoyl phorbol acetate (TPA) and cholera toxin. We also derived an immortal line of pigmented melanocytes from the epidermis of a healthy Meishan pig. This cell line, designated PigMel, retains differentiation function in culture, dependence on TPA and cholera toxin and a diploid chromosome number. PigMel melanocytes exhibit morphological and molecular characteristics common to normal mammalian skin melanocytes. [source] Expression of Notch signalling-related genes in normal and differentiating rat dental pulp cellsAUSTRALIAN ENDODONTIC JOURNAL, Issue 2 2010Hantang Sun dds Abstract Notch signalling is of fundamental importance to various processes during embryonic development and in adults. The possible role of Hey1, an important Notch signalling component, in odontoblast differentiation was evaluated in this study. Primary cultured dental pulp cells, derived from upper incisors of 5-week-old Wistar rats, were placed in ,-modification of Eagle's minimal essential medium supplemented with 10% Fetal Bovine Serum (FBS), and ascorbic acid (AA) and ,-glycerophosphate (,-GP), with or without dexamethasone, and cultured on dishes coated with collagen type IA for 7 days. Conventional and real-time Polymerase Chain Reaction (PCR) was performed to determine the expression of Notch-related genes and dentin sialophosphoprotein as a marker of odontoblast differentiation. Dentin sialophosphoprotein and Hey1 expression was significantly increased and decreased in the presence of AA + ,-GP compared with controls, respectively. These findings suggest that Hey1 may be a negative regulator in odontoblast differentiation. [source] | ||||||||||