A simple method for preparing single cell suspensions of heart and smooth muscle for radioreceptor labeling studies.
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.
D000818:Animals; D005260:Female; D006168:Guinea Pigs; D009130:Muscle, Smooth / Q000032:analysis*; D009206:Myocardium / Q000032:analysis*; D011813:Quinuclidinyl Benzilate / Q000378:metabolism; D011869:Radioligand Assay / Q000379:methods*; D011976:Receptors, Muscarinic / Q000032:analysis
D011976:Receptors, Muscarinic; D011813:Quinuclidinyl Benzilate