Preparation of gonadotroph-enriched cell populations from adult rat anterior pituitary cells by centrifugal elutriation.
Preparation of gonadotroph-enriched cell populations from adult rat anterior pituitary cells by centrifugal elutriation.
复制标题
通过离心淘析从成年大鼠垂体前叶细胞中制备富含促性腺激素的细胞群。
作者：
C. Hyde;G. Childs;L. Wahl;Z. Naor;K. Catt
采用离心洗脱法从成年雌性大鼠垂体细胞悬浮液中分离促性腺激素，制备促性腺激素富集细胞。在转子转速为1960 rpm时，将胰蛋白酶分散的细胞（2 - 3亿个）送入洗脱室，通过增加通过转子的介质流速，从11.8 ml/min增加到39.5 ml/min，将细胞进行分段分离和收集。80 min内完成上样和洗脱；细胞回收率为78/95%。用亲和素-生物素-过氧化物酶复合物技术对初始细胞悬液（ICS）进行免疫细胞化学染色，结果显示8%的细胞含有LH-β， 12%的细胞含有FSH-β。洗脱后，在分数7中发现大的成熟的促性腺激素，以30.8/39.5 ml/min的速度收集。在这个部分中，40%的细胞被LH-β染色，47%的细胞被FSH- β染色。由于分数7中近一半的细胞是内皮细胞，促性腺激素至少占分泌量的80%。
Separation of gonadotrophs from cell suspensions of adult female rat pituitary glands by centrifugal elutriation was applied to the preparation of gonadotropin-enriched cells. Trypsin-dispersed cells (200–300 million) were loaded into the elutriator chamber while the rotor was operating at 1960 rpm, and cells were separated and collected in fractions by increasing the flow rate of medium through the rotor in increments from 11.8 to 39.5 ml/min. Loading and elutriation were completed in 80 min; cell recovery was 78/95%. Immunocytochemical staining of the initial cell suspension (ICS) by the avidin-biotin-peroxidase complex technique demonstrated that 8% of the cells contained LH-β and 12% contained FSH-β. After elutriation, large, mature gonadotrophs were found in fraction 7, collected at 30.8/39.5 ml/min. In this fraction, 40% of the cells stained for LH-β and 47% stained for FSH-(β. Since nearly half of the cells in fraction 7 were endothelial cells, the gonadotrophs constituted at least 80% of the secre...
DOI：
10.1177/30.7.6286753
发表时间：
1982-07
期刊：
The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society
影响因子：
--
作者：
G. Childs;G. Unabia
通讯作者：
G. Childs;G. Unabia
应用快速亲和素-生物素-过氧化物酶复合物（ABC）技术在电子显微镜水平上定位垂体激素。
DOI：
10.1177/30.12.6296223
发表时间：
1982
期刊：
The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society
影响因子：
--
作者：
Childs,GV;Unabia,G
通讯作者：
Unabia,G