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A serum-free culture system for stydying solute exchanges in the choroid plexus
Authors:W F Agnew  R B Alvarez  T G H Yuen  S B Abramson  D Kirk
Institution:(1) Neurological Research Laboratory, Huntington Medical Research Institutes, 91105 Pasadena, California
Abstract:Summary Organ cultures of choroid plexus tissues from the lateral ventricle of juvenile rats have been maintained for periods up to 7 wk in a chemically defined, serum-free media. Of several media and various supplements evaluated, the best growth and survival was obtained with the Pasadena Foundation for Medical Research-4 media supplemented with three hormones: epidermal growth factor, insulin, and hydrocortisone. Autoradiographic studies demonstrated that the epithelial cells incorporated 3H]leucine and 3H]thymidine indicating active protein and DNA synthesis, respectively. The organ cultures were characterized by bulbous, vesicular outgrowths from the choroidal villi explants. The fluid-filled lumina of the vesccles reached diameters of 900 μm and were easily accessed by micropipettes. The walls of the vesicles were composed of single layers of epithelial cells in which the ultrastructural features in the in vivo tissue were well maintained. The in vivo polarity (apical end toward the media and basilar end of the cells toward the luminal cavity) was also maintained. This morphologically stable in vitro system seems to be a promising model for investigation of secretory mechanisms of choroidal tissue. This work was supported in part by National Institutes of Health Grant NS 12906-06.
Keywords:choroid plexus  organ culture  serum-free cultures  model system  epithelium  rat
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