Notter M F, Gash D M, Sladek C D, Scharoun S L
Brain Res Bull. 1984 Mar;12(3):307-13. doi: 10.1016/0361-9230(84)90058-3.
A microsystem for rotation-mediated aggregate cell culture studies has been devised to examine vasopressin (VP) biosynthesis of developing rat hypothalamus. Trypsin-dispersed hypothalamic tissue was placed into 24 well tissue culture dishes and VP content of culture medium and cells was measured over time by a radioimmunoassay. Reaggregates formed within 4 hr when rotated at 70 rpm in a humid CO2 incubator. Nineteen days post coitus (dpc) hypothalamic reaggregates had 336 pg VP/10(6) cells while the medium showed 260 pg VP/ml after four days. Measurable VP was seen in fetal tissue after ten days while comparable amounts of VP were present in one day neonatal hypothalamus over this same period. Morphological examination of reaggregates indicated the presence of viable cells throughout the cell mass after ten days of culture. Co-cultivation studies with dispersed posterior pituitary indicated that reaggregates from one day neonate hypothalamus had significantly increased VP levels when co-cultured with one day neonatal posterior pituitary; however, this effect was not seen with 19 dpc co-cultures. These data demonstrate that development of neurosecretory activity of discrete regions of the hypothalamus can be examined early in vitro in a reaggregate cell culture system.
为研究发育中大鼠下丘脑的血管加压素（VP）生物合成，设计了一种用于旋转介导的聚集细胞培养研究的微系统。将胰蛋白酶分散的下丘脑组织置于24孔组织培养板中，并通过放射免疫测定法随时间测量培养基和细胞中的VP含量。在潮湿的二氧化碳培养箱中以70转/分钟旋转时，4小时内形成再聚集物。交配后19天（dpc）的下丘脑再聚集物每10^6个细胞含336 pg VP，而4天后培养基中VP含量为260 pg/ml。培养10天后在胎儿组织中可见可测量的VP，而同期1日龄新生鼠下丘脑VP含量与之相当。再聚集物的形态学检查表明，培养10天后整个细胞团中均存在活细胞。与分散的垂体后叶共培养研究表明，1日龄新生鼠下丘脑的再聚集物与1日龄新生鼠垂体后叶共培养时VP水平显著升高；然而，19 dpc共培养时未观察到这种效应。这些数据表明，下丘脑离散区域神经分泌活性的发育可在体外早期通过再聚集细胞培养系统进行研究。
