Johns A, Riehl R M
J Pharmacol Methods. 1982 Mar;7(2):153-9. doi: 10.1016/0160-5402(82)90028-6.
A simple method of preparing single cell suspensions for radioligand binding studies is described. The method involves incubating tissues in the presence of collagenase and elastase for 90 min in physiological solution with 1 mM calcium chloride and the mechanical disruption of the tissue by pipetting. The tissues examined were atria, ventricle, bladder, uterus, and taenia coli removed from mature guinea pigs. Viability of the cells by trypan blue exclusion showed 60-88% viable cells and receptor binding studies using (3H]-1-quinuclidinyl benzilate (QNB) yield KD values of approximately equal to 0.1 nM. The receptor numbers for each tissue were (receptors/cell): atria, 5700; ventricle, 11,000; uterus, 31,000; bladder, 44,000; taenia coli, 68,000.
本文描述了一种用于放射性配体结合研究的单细胞悬液制备的简单方法。该方法包括在含有1 mM氯化钙的生理溶液中，将组织与胶原酶和弹性蛋白酶一起孵育90分钟，然后通过移液管对组织进行机械破碎。所检查的组织取自成年豚鼠的心房、心室、膀胱、子宫和结肠带。经台盼蓝拒染法检测，细胞活力显示60 - 88%的细胞存活，使用[³H]-1-喹核醇基苯甲酸酯（QNB）进行的受体结合研究得出的KD值约为0.1 nM。每个组织的受体数量为（受体/细胞）：心房，5700；心室，11,000；子宫，31,000；膀胱，44,000；结肠带，68,000。
