A neural extracellular matrix-based method for in vitro hippocampal neuron culture and dopaminergic differentiation of neural stem cells

Background: The ability to recreate an optimal cellular microenvironment is critical to understand neuronal behavior and functionality in vitro. An organized neural extracellular matrix (nECM) promotes neural cell adhesion, proliferation and differentiation. Here, we expanded previous observations on the ability of nECM to support in vitro neuronal differentiation, with the following goals: (i) to recreate complex neuronal networks of embryonic rat hippocampal cells, and (ii) to achieve improved levels of dopaminergic differentiation of subventricular zone (SVZ) neural progenitor cells. Methods: Hippocampal cells from E18 rat embryos were seeded on PLL- and nECM-coated substrates. Neurosphere cultures were prepared from the SVZ of P4-P7 rat pups, and differentiation of neurospheres assayed on PLL- and nECM-coated substrates. Results: When seeded on nECM-coated substrates, both hippocampal cells and SVZ progenitor cells showed neural expression patterns that were similar to their poly-L-lysine-seeded counterparts. However, nECM-based cultures of both hippocampal neurons and SVZ progenitor cells could be maintained for longer times as compared to poly-L-lysine-based cultures. As a result, nECM-based cultures gave rise to a more branched neurite arborization of hippocampal neurons. Interestingly, the prolonged differentiation time of SVZ progenitor cells in nECM allowed us to obtain a purer population of dopaminergic neurons. Conclusions: We conclude that nECM-based coating is an efficient substrate to culture neural cells at different stages of differentiation. In addition, neural ECM-coated substrates increased neuronal survival and neuronal differentiation efficiency as compared to cationic polymers such as poly-L-lysine.

Potentzia-bihurgailu matrizialen diseinu-prozesurako eta hutsegite-tolerantziarako soluzio berriak

290 p.

Linear Chromatic Adaptation Transform Based on Delaunay Triangulation

Computer vision algorithms that use color information require color constant images to operate correctly. Color constancy of the images is usually achieved in two steps: first the illuminant is detected and then image is transformed with the chromatic adaptation transform ( CAT). Existing CAT methods use a single transformation matrix for all the colors of the input image. The method proposed in this paper requires multiple corresponding color pairs between source and target illuminants given by patches of the Macbeth color checker. It uses Delaunay triangulation to divide the color gamut of the input image into small triangles. Each color of the input image is associated with the triangle containing the color point and transformed with a full linear model associated with the triangle. Full linear model is used because diagonal models are known to be inaccurate if channel color matching functions do not have narrow peaks. Objective evaluation showed that the proposed method outperforms existing CAT methods by more than 21%; that is, it performs statistically significantly better than other existing methods.

Linear Chromatic Adaptation Transform Based on Delaunay Triangulation

Computer vision algorithms that use color information require color constant images to operate correctly. Color constancy of the images is usually achieved in two steps: first the illuminant is detected and then image is transformed with the chromatic adaptation transform ( CAT). Existing CAT methods use a single transformation matrix for all the colors of the input image. The method proposed in this paper requires multiple corresponding color pairs between source and target illuminants given by patches of the Macbeth color checker. It uses Delaunay triangulation to divide the color gamut of the input image into small triangles. Each color of the input image is associated with the triangle containing the color point and transformed with a full linear model associated with the triangle. Full linear model is used because diagonal models are known to be inaccurate if channel color matching functions do not have narrow peaks. Objective evaluation showed that the proposed method outperforms existing CAT methods by more than 21%; that is, it performs statistically significantly better than other existing methods.