Human embryonic stem cells (hESCs) are expected to open up new avenues in regenerative medicine by allowing the generation of transplantable cells to be used in future cell replacement therapies. Maintenance of hESCs in the presence of xenogenic compounds is likely to prevent their use in future therapeutic applications in humans. Recently, it has been claimed that human foreskin-derived human embryonic fibroblast (HEFs) and human adult marrow cells have the ability to support prolonged expansion of hESCs in culture similar to murine feeders. Here, to minimize the use of xenogenic components for hESC maintenance, we performed transmission electron microscopy-based microbiological studies in an attempt to implement a microbiological Quality Assurance Program in Stem Cell Banks by determining the potential presence of viral particles in MEFs compared with human HEFs and bone marrow-derived mesenchymal cells. We observed in three out of nine MEF samples (33.3%) viruses belonging to the Retroviridae family. Within the Retroviridae family, these viruses have a C morphology, which indicates they belong to the subfamily Orthoretroviridae. In contrast, no viral particles could be observed in either the HEF samples (n = 5) or the human BM-derived mesenchymal cells (n = 9) analyzed. Based on these experimental microbiological data, we recommend the implementation of microbiological Quality Assurance Programs by means of transmission electron microscopy as a routine technique to assess the potential presence of viral particles in any feeder cell used in stem cell banks and support the use of human cells rather than murine cells as feeders to maintain hESC cultures in an undifferentiated state.

Purpose: This paper aims to study the anterior surface of the optic nerve in relation to its ability to support a source of stress acting from the vitreous cavity. The intercellular junctions of the lining astrocytes mediated by cellular adhesion molecules (CAMs) may be the main targets for ionic stress. Methods: The optic nerve of the domestic pig was prepared for light, confocal laser and transmission electron microscopy. Immunostaining was performed for antibodies against glial fibrillary acidic protein, neural cadherin (N-cadherin) and neural CAM (N-CAM). Results: Only 1 type of intercellular junction was found among the bordering astrocytes, which was characterized as a zonula adherens. Unions between lining cells showed a positive immunogold effect and immunofluorescence against N-cadherin in the zonula adherens and membrane apposition. N-CAM was also present in areas of nonjunctional cellular adhesion. Conclusion: The stability of intercellular junctions of the nerve-vitreous boundary is sensitive to altered concentrations of Ca²⁺. Since aqueous humor has half the Ca²⁺ concentration of plasma, any contact of aqueous humor with the optic nerve head can interfere with the ionic concentration of calcium in the extracellular spaces. This mechanism may contribute to age-related changes and some types of glaucoma.

A thin mucous layer covers the ciliary body and iris epithelium and becomes thicker in the anterior chamber where it covers the anterior surface of the iris, the chamber angle and corneal endothelial surface. It is especially thick at the chamber angle, where it adopts a meniscus shape with the concavity towards the anterior chamber. Cetylpiridinium chloride along with glutaraldehyde was used to precipitate this layer of mucous substance lining the anterior and posterior chambers of the human eye. The staining of the precipitate by cationic dyes suggests that glycosaminoglycans are main components. The specific characterisation with anti-hyaluronic acid monoclonal antibody labelled with colloidal gold reveals the presence of hyaluronic acid in the precipitate. Long unbranched hyaluronic acid molecules may form the skeleton of a gel able to trap and hold virtually any other macromolecule suspended in the aqueous humour. A possible role of such a gel in the regulation of aqueous humour outflow is discussed.

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Delineation of the astrocytic pavement and the preferred flow routes formed by wide extracellular spaces are conspicuous features of the prelaminar region when examined with the confocal laser microscope and the help of sticky fluorescent tracer. This suggests that excess extracellular fluid can be interchanged with the vitreous by a network of interconnected extracellular spaces or preferred flow routes. Some pathogenic mechanisms can be related to fluid interchange in the optic nerve head.