研究脉络膜丛溶质交换的无血清培养系统。

In Vitro Pub Date : 1984-09-01 DOI:10.1007/BF02618877
W F Agnew, R B Alvarez, T G Yuen, S B Abramson, D Kirk
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引用次数: 3

摘要

幼年大鼠侧脑室脉络膜丛组织的器官培养在化学定义的无血清培养基中维持了长达7周。在评估的几种培养基和各种补充剂中,帕萨迪纳医学研究基金会-4培养基中添加三种激素:表皮生长因子、胰岛素和氢化可的松,获得了最佳的生长和存活率。放射自显影研究表明,上皮细胞分别掺入[3H]亮氨酸和[3H]胸苷,表明活性蛋白和DNA合成。器官培养的特点是由脉络膜绒毛外植体生长出球根状、水疱状的外植体。囊泡内充满液体的腔直径达到900微米,用微移液器很容易接近。囊泡的壁由单层上皮细胞组成,其体内组织的超微结构特征保持得很好。细胞的体内极性(细胞的根尖端朝向介质,细胞的基底端朝向管腔)也保持不变。这种形态稳定的体外系统似乎是研究脉络膜组织分泌机制的一个有前途的模型。
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A serum-free culture system for studying solute exchanges in the choroid plexus.

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 vesicles reached diameters of 900 microns 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.

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