An in Vitro Model for Arsine Toxicity Using Isolated Red Blood Cells

Abstract

An in Vitro Model for Arsine Toxicity Using Isolated Red Blood Cells. Hatlelid, K. M., Brailsford, C., and Carter, D. E. (1995). Fundam. Appl. Toxicol. 25, 302-306. A novel test system using isolated red blood cells (RBCs) and arsine (AsH 3) in aqueous solution was developed to allow quantitation of AsH 3 exposure and to study the toxicity of AsH 3 in vitro. In this system AsH 3 gas was generated and dissolved in aqueous solution, the concentration was measured, and the standardized solution was mixed with rat or dog red blood cells (RBCs). AsH 3 was found to be stable in solution at neutral pH for several hours, but was lost quickly from solution as the acidity was increased to pH 2. Approximately 74% of the initial 0.56 mM AsH 3, measured as total arsenic, was found to be taken up by, or strongly associated with, dog RBCs within 5 min of incubation and 82% of the initial 0.49 mM AsH 3 was found in rat RBCs after 10 min incubation. Following hypotonic lysis of rat RBCs, 55% of the cell-associated arsenic was found in the membrane fraction with the balance found in the cytosolic fraction. The in vitro technique was used to examine factors influencing AsH 3 toxicity using hemolysis as the end point. Hemolysis levels in dog and rat RBC incubations were found to increase with time after exhibiting a lag phase of about 30 min. At the AsH 3 concentrations used, maximum levels of hemolysis were observed by 2 hr; maximum hemolysis at room temperature for dog RBCs was 20% and for rat RBCs was 22%. Increasing the temperature from room temperature to 37°C resulted in increased hemolysis in dog RBCs (36%) and rat RBCs (90%). Increasing the concentration of AsH 3 in the incubations also resulted in increased hemolysis; approximately 9% hemolysis was observed after 2 hr incubation of rat RBCs with 0.12 mM AsH 3 while 30% hemolysis was observed following incubation with 0.59 mM AsH 3. After this in vitro system is characterized, it will be used to investigate theories of AsH 3 toxicity put forth in the literature.