Agnew W F, Alvarez R B, Yuen T G, Abramson S B, Kirk D
In Vitro. 1984 Sep;20(9):712-22. doi: 10.1007/BF02618877.
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.
来自幼年大鼠侧脑室脉络丛组织的器官培养物,已在化学成分明确的无血清培养基中维持长达7周。在评估的几种培养基和各种补充剂中,添加三种激素(表皮生长因子、胰岛素和氢化可的松)的帕萨迪纳医学研究基金会-4培养基能实现最佳的生长和存活效果。放射自显影研究表明,上皮细胞分别掺入了[3H]亮氨酸和[3H]胸腺嘧啶核苷,这分别表明有活跃的蛋白质和DNA合成。器官培养物的特征是脉络丛绒毛外植体上有球状、囊状的生长物。囊泡中充满液体的腔直径可达900微米,很容易用微量移液器进入。囊泡壁由单层上皮细胞组成,其中体内组织的超微结构特征得到了很好的维持。体内的极性(细胞的顶端朝向培养基,基底端朝向管腔)也得以维持。这个形态学上稳定的体外系统似乎是研究脉络丛组织分泌机制的一个有前景的模型。