A multi-tissue segmentation of the human head for detailed computational models.

This paper describes the creation of an anatomically detailed high resolution model of the human head based on the Visible Human Female data from the National Library of Medicine archives. Automatic and semi-automatic segmentation algorithms were applied over the 3 image volumes ­ CT, MRI and anatomical cryo-sections of the cadaver ­ to label a total of 23 tissues. The results were combined to create a labeled volume of the head with voxel dimensions of 0.33×0.33×0.33 mm. The individual label matrices and their corresponding surface meshes are made available to be used freely. The detailed blood vessel network and ocular tissues will be of interest in computational modelling and simulation studies.

Impedance spectra of polypyrrole coated platinum electrodes.

Polypyrrole (PPy) coated electrodes may provide new solutions to increase the charge injection capacity and biocompatibility of metal electrodes in e.g., neural stimulus applications. In this study, electrical impedance spectra of PPy coated platinum (Pt) electrodes having three different coating thicknesses were measured and modeled. A suitable equivalent electrical circuit providing the material characteristics was chosen and the impedance data was analyzed using the model and data fitting. The modeled parameter values of different coating thicknesses were compared and our results demonstrated the changes in charge transfer properties and mechanisms of thin and thick PPy film coatings.

In vitro electroretinogram for the study of the functionality of differentiated retinal pigment epithelium cells.

The purpose of this study is to develop and test a method to reveal if the retinal pigment epithelium (RPE) cells differentiated from human embryonic stem cells (hESC) support the functions of photoreceptors. hESC-derived RPE (hESC-RPE) cells offer a potent cell source for cell replacement therapy that may be used to prevent certain eye diseases. Methods to assure the functionality of the RPE cells are well warranted. Electroretinograms (ERG) measure the electrophysiological response of the retina to light stimuli. A setup was developed that enables the measurement of ERG in vitro from mice retinas cultured together with hESC-RPE cells. The co-culture of RPE and retinas seems to be a viable tool to assess the functionality of RPE in vitro. However, owing to limited sample size results were somewhat mixed, and thus it was not possible to prove that hESC-RPE cells enhance the ERG response of a mouse retina in vitro. The long-term culturing of the retinas needs to be refined to acquire more conclusive evidence of the supporting role of the RPE and to explore the full potential of the co-culture and ERG methods in assessing RPE functionality.

Electric impedance of human embryonic stem cell-derived retinal pigment epithelium.

The barrier properties of epithelium are conventionally defined by transepithelial resistance (TER). TER provides information about the tightness of the epithelium. Electrical impedance spectroscopy (EIS) provides additional information regarding cell membrane properties, such as changes in electric capacitance and possible parallel or serial pathways that may correlate with the morphology of the cell layer. This study presents EIS of retinal pigment epithelial (RPE) cell model of the putative RPE differentiated from human embryonic stem cells (hESC-RPE). The generally utilized RPE cell model, ARPE-19, was used as immature control. The measured EIS was analyzed by fitting an equivalent electrical circuit model describing the resistive and capacitive properties of the RPE. Our results indicated that TER of hESC-RPE cells was close to the values of human RPE presented in the literature. This provides evidence that the stem cell-derived RPE in vitro can reach high-barrier function. Furthermore, hESC-RPE cells produced impedance spectra that can be modeled by the equivalent circuit of one time constant. ARPE-19 cells produced low-barrier properties, that is, an impedance spectra that suggested poor maturation of ARPE-19 cells. To conclude, EIS could give us means for non-invasively estimating the functionality and maturation of differentiated-RPE cells.

Comparison of electrode materials for the use of retinal prosthesis.

Retinal implants may provide vision for people suffering from photoreceptor degeneration caused by different eye diseases. Electrode size in retinal implant should be decreased in order to increase the resolution provided by the implant. We defined electric properties of five different electrode materials (Au, Ir-b, Ti, TiN, Pt-b) widely used in retinal prostheses. The comparison of different electrode materials requires that the electrical properties of different materials are defined using exactly the same measurement conditions and devices. Existing studies about electrode material properties are often made using slightly different measurement parameters or electrode processing conditions making the comparison between different materials difficult. Here, the electrochemical characterization included cyclic voltammetry and electrochemical impedance spectroscopy. Ir-b and Pt-b had greater charge injection capacity than other materials. The fabricated material samples showed that in this experiment the electrode diameter larger than 200 µm should be used to suppress irreversible reaction of stimulus electrodes with the needed stimulus currents. Thus, either we have to find novel electrode materials or surface treatment methods to decrease the electrode area providing increased electrode and pixel number of the prosthesis or we have to show that stimulus currents smaller than 40 µA are enough to induce phosphenes.

Toward the defined and xeno-free differentiation of functional human pluripotent stem cell-derived retinal pigment epithelial cells.

PURPOSE: The production of functional retinal pigment epithelium (RPE) cells from human embryonic (hESCs) and human induced pluripotent stem cells (hiPSCs) in defined and xeno-free conditions is highly desirable, especially for their use in cell therapy for retinal diseases. In addition, differentiated RPE cells provide an individualized disease model and drug discovery tool. In this study, we report the differentiation of functional RPE-like cells from several hESC lines and one hiPSC line in culture conditions, enabling easy translation to clinical quality cell production under Good Manufacturing Practice regulations. METHODS: Pluripotent stem cells were cultured on human fibroblast feeder cells in serum-free medium. The differentiation toward RPE was induced by removing basic fibroblast growth factor and feeder cells from the serum-free conditions. RPE differentiation was also achieved using xeno-free and defined culture conditions. The RPE cell morphology and pigmentation of the cells were analyzed and the expression of genes and proteins characteristic for RPE cells was evaluated. In vitro functionality of the cells was analyzed using ELISA measurements for pigment epithelium derived factor (PEDF) secretion and phagocytosis of photoreceptor outer segments (POS). The integrity of the generated RPE layers was analyzed using transepithelial electric resistance measurements. RESULTS: We generated putative RPE cells with typical pigmented cobblestone-like morphology. The expression of RPE-specific markers was confirmed at the gene and protein level. The differentiated cells were able to phagocytose POS and secrete PEDF characteristic of native RPE cells. In addition, cultured cells formed a polarized epithelium with high integrity and exhibited excellent transepithelial electric resistance values, indicating well established, tight junctions. Moreover, we introduced an improved method to generate functional putative RPE cells without xeno-components under defined conditions. CONCLUSIONS: We have developed a progressive differentiation protocol for the production of functional RPE-like cells from hESCs and hiPSCs. Our results demonstrate that putative hESC-RPE and hiPSC-RPE express genes and proteins characteristic for RPE cells, as well as being able to phagocytose POS and secrete PEDF. Furthermore, our results show that RPE-like cells can be differentiated in xeno-free and defined culture conditions, which is mandatory for Good Manufacturing Practice-production of these cells for clinical use.