Measurement of red cell sickling: a method for studying the efficacy of antisickling drugs under physiological conditions.
A method was developed to study sickling in vitro under physiological conditions using a small amount of blood (0.1 mL). The diluted blood suspension (2.1 mL) was placed in a flask and flushed with a gas mixture containing 5% CO2. In deoxygenation experiments, samples were withdrawn anaerobically into a microslide (optical path 0.1 mm) and red cell morphology was studied directly under a light microscope after both ends of the microslide were sealed. The blood suspension with a hematocrit value of 1% can be deoxygenated in less than 10 min, but it takes 30 min for the sickling of cells to reach a plateau. The degree of sickling increases with increasing osmolality of the medium, or with a decrease of the pH. With a citrate-phosphate-dextrose-adenine solution, sickle blood may be stored for this type of study for about 10 days at 4 degrees C. The blood may be stored for about 5 days with a noncitrate preservative. This method was found useful in examining the antisickling activity of various drugs.
D000328:Adult; D000755:Anemia, Sickle Cell / Q000473:pathology*; D000986:Antisickling Agents / Q000627:therapeutic use*; D001793:Blood Preservation; D004341:Drug Evaluation; D004912:Erythrocytes / Q000473:pathology*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D009994:Osmolar Concentration; D012805:Sickle Cell Trait / Q000188:drug therapy / Q000473:pathology*; D013048:Specimen Handling; D013696:Temperature; D013997:Time Factors
D000986:Antisickling Agents