No abstract available.
Calcium Signaling/physiology* ; Cell Polarity/physiology* ; Epithelial Cells/physiology* ; Epithelial Cells/cytology*

The aim of this study was to isolate, cultivate and phenotypically characterize two types of human amnio-tic membrane (HAM)-derived cells, and to analyze their differentiation potential in vitro. Human amnion epithelial cells (hAEC) were derived from the embryonic ectoderm, while human amnion mesenchymal cells (hAMC) were derived from the embryonic mesoderm. The cells were characterized by flow cytometry and immunofluorescence, then immunofluorescence also was performed for the analysis of multipotentiality in differentiation. The results indicated that immunophenotypic characterization of both cell types demonstrated positive for HLA-A, B, C and mesenchymal stem cell markers (CD29, CD73, CD44, CD59, CD90, CD105, CD166), but did not express the hematopoietic markers (CD31, CD34, CD45, HLA-DR) and showed the weak expression of costimulatory molecules (CD40, CD40L, CD80, CD86). Phenotypes of both cell populations were maintained from passages 3 to 7. The immunofluorescence indicated that hAEC expressed cytokeratin 19, but did not express vimentin. On the contrary, hAMC expressed vimentin but did not express cytokeratin 19. The assessment of multilineage potential demonstrated that hAMC showed greater cardiomyocytes potential, while hAEC showed greater neural potential. It is concluded that hAEC and hAMC can be successfully isolated from the HAM. Both cell populations possess similar immunophenotype. However, they differ in cell yield and multipotential for differentiation into the major lineages, hAEC possess a much greater ectodermal differentiation capacity, while hAMC possess a much greater mesodermal differentiation capacity. This conclusion will be important for use of these cells in cell therapy.
Amnion ; cytology ; Cell Differentiation ; physiology ; Cell Lineage ; Epithelial Cells ; cytology ; Humans ; Immunophenotyping ; Stromal Cells ; cytology

Mesothelial cells (MCs) cover the surface of visceral organs and the parietal walls of cavities, and they synthesize lubricating fluids to create a slippery surface that facilitates movement between organs without friction. Recent studies have indicated that MCs play active roles in liver development, fibrosis, and regeneration. During liver development, the mesoderm produces MCs that form a single epithelial layer of the mesothelium. MCs exhibit an intermediate phenotype between epithelial cells and mesenchymal cells. Lineage tracing studies have indicated that during liver development, MCs act as mesenchymal progenitor cells that produce hepatic stellate cells, fibroblasts around blood vessels, and smooth muscle cells. Upon liver injury, MCs migrate inward from the liver surface and produce hepatic stellate cells or myofibroblast depending on the etiology, suggesting that MCs are the source of myofibroblasts in capsular fibrosis. Similar to the activation of hepatic stellate cells, transforming growth factor β induces the conversion of MCs into myofibroblasts. Further elucidation of the biological and molecular changes involved in MC activation and fibrogenesis will contribute to the development of novel approaches for the prevention and therapy of liver fibrosis.
Epithelial Cells/*physiology ; Epithelium/metabolism ; Hepatic Stellate Cells/*physiology ; Humans ; Liver/*cytology/injuries/*physiology ; Liver Cirrhosis/etiology/prevention & control ; Liver Regeneration/*physiology ; Mesenchymal Stromal Cells/physiology ; Myofibroblasts/physiology

OBJECTIVETo investigate the effects of ephedrine on human nasal cilia movement.

METHODSCiliary beat frequency (CBF) of cultured human nasal epithelial cells was measured by high-speed digital microscopy in HBSS and ephedrine solution of different concentrations in 10 minutes.

RESULTSCBF of cultured nasal epithelial cells exposed to HBSS showed no significant changes in 10 minutes. However, in 2.5 g/L , 5 g/L, 10 g/L and 20 g/L ephedrine solution, CBF increased significantly in 1-2 minutes and reached the apex, then it decreased gradually, at the 10th minute. CBF of the samples exposed to 2.5 g/L and 5 g/L ephedrine solution were slower than those in HBSS, but no significant changes were found. However, in 10 g/L and 20 g/L ephedrine solution, CBF decreased significantly when compared with samples in sHBSS. With the concentrations from 2.5 g/L to 20 g/L ephedrine, the increment was independent on the concentration, the inhibitory effect was dependent on the concentration.

CONCLUSIONSIn initial time, 2. 5 g/L-20 g/L ephedrine stimulated CBF, then 10 g/L-20 g/L ephedrine inhibited CBF. The stimulation of 2.5 g/L and 5 g/L ephedrine on CBF was longer than that of 10 g/L and 20 g/L ephedrine. 5 g/L ephedrine had maximum stimulatory effect without obvious inhibitory effect on cultured human nasal CBF.

Cells, Cultured ; Cilia ; drug effects ; physiology ; Ephedrine ; pharmacology ; Epithelial Cells ; drug effects ; physiology ; Humans ; Nasal Mucosa ; cytology ; drug effects ; physiology

Alveolar Epithelial Cells ; metabolism ; pathology ; Animals ; Apoptosis ; physiology ; Cell Survival ; physiology ; Cells, Cultured ; Epithelial Cells ; cytology ; physiology ; Genes, bcl-2 ; genetics ; Genes, p53 ; genetics ; Oxidative Stress ; genetics ; Rats

The Caco-2 cell model established as a tool for in vitro investigations of intestinal drug transport processes has been widely used because of its growth characteristics, i.e., it forms polarized monolayers in cultures and differentiates into cells with high homology to human intestinal epithelial absorptive cells. Caco-2 cell cultures have provided a major conceptual advance in our understanding of intestinal drug absorption, biotransformation and bioavailability at the cellular level. Caco-2 cells have received considerable attention from the pharmaceutical industry because they have been widely accepted as a potent in vitro model membrane to screen for potential absorption problems in drug discovery programs. However, the Caco-2 monolayers model is still not perfect. The tightness of the monolayers resembles more colonic than small intestinal tissue, resulting in poor permeabilities for hydrophilic compounds traversing the epithelium via the aqueous paracellular pathway. Caco-2 cells have no mucus layer that is a potential barrier to drug absorption and display low expression of cytochrome P450 which are drug metabolizing enzymes. Further refinements of the Caco-2 cell culture model are needed to better predict human intestinal drug transport. To optimize Caco-2 model, the following technics have been used: modifying the condition of the cell culture, using molecular cloning strategies and inducing the expression of relevant enzymes. They are described in this review.
Biological Availability ; Biological Transport ; Caco-2 Cells ; cytology ; Colon ; physiology ; Epithelial Cells ; cytology ; Humans ; Intestinal Absorption ; Models, Biological

The vibrational spectral differences of normal and lung cancer cells were studied for the development of effective cancer cell screening by means of attenuated total reflection infrared spectroscopy. The phosphate monoester symmetric stretching nus(PO3(2-)) band intensity at ~970 cm-1 and the phosphodiester symmetric stretching nus(PO2-) band intensity at ~1,085 cm-1 in nucleic acids and phospholipids appeared to be significantly strengthened in lung cancer cells with respect to the other vibrational bands compared to normal cells. This finding suggests that more extensive phosphorylation occur in cancer cells. These results demonstrate that lung cancer cells may be prescreened using infrared spectroscopy tools.
*Carcinoma ; Cell Line, Tumor ; Epithelial Cells/*physiology ; Humans ; *Lung Neoplasms ; Respiratory Mucosa/*cytology ; *Spectrophotometry, Infrared

The vectorial transepithelial transport of water and electrolytes in the renal epithelium is achieved by the polarized distribution of various transport proteins in the apical and basolateral membrane. The short-term regulation of transepithelial transport has been traditionally thought to be mediated by kinetic alterations of transporter without changing the number of transporters. However, a growing body of recent evidence supports the possibility that the stimulus-dependent recycling of transporter-carrying vesicles can alter the abundance of transporters in the plasma membrane in parallel changes in transepithelial transport functions. The abundance of transporters in the plasma membrane is determined by net balance between stimulus-dependent exocytic insertion of transporters into and endocytic retrieval of them from the plasma membrane. The vesicular recycling occurs along the tracts of the actin microfilaments and microtubules with associated motors. This review is to highlight the importance of vesicular transport in the short-term regulatory process of transepithelial transport in the renal epithelium. In the short-term regulation of many other renal transporters, vesicular transport is likely to be also involved. Thus, vesicular transport is now emerged as a wide-spread general regulatory mechanism involved in short-term regulation of renal functions.
Animal ; Biological Transport/physiology ; Endocytosis/physiology* ; Epithelial Cells/enzymology* ; Epithelial Cells/cytology* ; Exocytosis/physiol(HCMV)* ; H(+)-Transporting ATP Synthase/metabolism* ; Human ; Sodium Channels/metabolism

AIMTo establish the whole-cell recording techniques of the neuronal alpha4beta2, alpha4beta4, and alpha7-nicotinic acetylcholine receptors heterologously expressed in SH-EP1 cell line and discuss the electrophysiological characteristics of their open states.

METHODSThe cells were cultured with DEME medium(high glucose) and suitable for electrophysiological experiments three days after passage. The receptors were induced from resting states into open states by rapid application of nicotine (alpha4beta2, alpha4beta4) or choline (alpha7).

RESULTSThe SH-EP1 cells cultured by this method were in good conditions and expressed plenty of receptors. Alpha4beta2, alph4beta4 and alpha7 inward currents could be induced by rapid application of agonists but had different dynamic processes against time. All the three types of currents were dose and voltage-dependent and had inward rectification property.

CONCLUSIONThe open states of neuronal alpha4beta2, alpha4beta4, and alpha7-nicotinic acetylcholine receptors and their transitions have distinct characteristics and the inward currents of all this three types of receptors are dose and voltage-dependent and have inward rectification property.

Brain ; cytology ; metabolism ; Cell Line ; Epithelial Cells ; cytology ; Humans ; Membrane Potentials ; physiology ; Neurons ; cytology ; metabolism ; Patch-Clamp Techniques ; Receptors, Nicotinic ; physiology ; Transfection ; alpha7 Nicotinic Acetylcholine Receptor

OBJECTIVESTo investigate the effects of newborn bull serum(NBS), vitamin C and vitamin E on cryopreservation of mouse seminiferous epithelial cells.

METHODSThe seminiferous epithelial cells from 7-day-old mice were cryopreserved in different freezing solutions. The cell recoveries were examined by Trypan blue exclusive staining after thawing. The freezing solutions composed of DMEM, 10% dimethylsulphoxide(DMSO), and 0, 5%, 10%, or 20% NBS, respectively, or composed of DMEM, 10% DMSO, 10% NBS, and 150 micrograms/ml vitamin C or 50 micrograms/ml vitamin E, respectively.

RESULTSThe cell recoveries in freezing solution containing 0, 5%, 10%, or 20% NBS were 83.4%, 84.7%, 85.7% and 83.6%, respectively. There were no significant differences between them. The cell recoveries in freezing solution containing vitamin C or vitamin E were 88.0% and 82.9%, respectively. There was no significant differences compared with that in freezing solution containing 10% DMSO and 10% NBS.

CONCLUSIONSNBS, vitamin C and vitamin E have no significant protecting effects on mouse seminiferous epithelial cells, and can not significantly improve the cell recoveries.

Animals ; Ascorbic Acid ; pharmacology ; Cattle ; Cryopreservation ; Epithelial Cells ; physiology ; Fetal Blood ; physiology ; Male ; Mice ; Seminiferous Epithelium ; cytology ; Vitamin K ; pharmacology