Blood cell lysis

Hemoglobinuria and hematin casts in the distal convoluted tubules and tubular lumens located in the medulla and papilla were reported in rats after a single dermal exposure to 107.1 mg/kg liquid phenol (Conning and Hayes 1970). These phenomena are probably related to red blood cell lysis and increased glomerular filtration of hemoglobin. Hemoglobinuria is characteristic of lethal or near-lethal exposures by the dermal route. 

The serum should be yellowish. A reddish color is indicative of erythrocytes lysis (hemolysis) which may interfere with clinical chemistry assays based on colorimetric values. If this happens, you may need to adjust the speed at which the blood is collected and processed or other steps that may cause sheer and red blood cell lysis. 

Red blood cells were lysed with NH4C1 red blood cell lysis buffer (pH 7.4). 

Chow S, Hedley D, Grom P, Magari R, Jacobberger JW, Shankey TV. Whole blood fixation and permeabilization protocol with red blood cell lysis for flow cytometry of intracellular phosphorylated epitopes in leukocyte subpopulations. Cytometry A 2005 67(1) 4-17. 

Prepcaration of Leucocytes from Peripheral Blood by Red Blood Cell Lysis (2 )... 

Inhibition of sheep red blood cell lysis by serum from sensitized mice is an indication of suppression of the humoral immune response. Groups of mice are sensitized by the intravenous injection of 0.2 ml of a 2% sheep red blood cell suspension. The test drug or vehicle is administered intraperitoneally to groups of mice for 3 consecutive days beginning 2h after... 

Red blood cell lysis buffer (ACK lysing buffer from BioSource International, Camarillo, CA) 156 mM NH4CI, 10 xaM KHCO3, and 1 mAf Na2EDTA. 

Exposure of human red blood cells from healthy adult volunteers of either sex to high concentration of HOCl and HOBr (>40 nmol/10 cells) resulted in rapid cell lysis and the detection of an additional nitrogen-centred, protein-derived radical adduct (Hawkins et al. 2001). HOBr induced red blood cell lysis at approximately 10-fold lower concentration than HOCl, whereas with monocyte (HTPl) and macrophage (J774) cells HOCl and HOBr induced lysis at similar concentrations. Erythrocytes exposed to nonlytic doses of HOCl generated novel nitrogen-centred radicals the formation of which is GSH dependent. 

Eliminate red blood cells (if necessary) by adding 10 ml of red blood cell lysis buffer. Pipette up and down. Leave cell suspension at room temperature for 5 min (debris and clumps of cells with sediment at the bottom of the tube). 

Fig. 2 Biocompatibility assays of polyacetal 16 described in Scheme 5. (a) Cytotoxicity assay (B16F10 cells stimulated for 72 h) positive control was poly-L-lysine, negative control was Dextran. (b) Red blood cell lysis assay after 24 h positive control was poly(ethylene imine) (PEI), negative control was Dextran. Adapted with permission from [53], Copyright 2002 American Chemical Society...

To investigate the safety of thiolated chitosans, several studies have been performed measuring its potential cytotoxicity on various cell lines. The toxicity profiles of chitosan-TBA, for example, were assessed using the red blood cell lysis test. The results showed that thiolated chitosan had a lower membrane damaging effect than the corresponding unmodified control. The safety of thiolated chitosan was further confirmed by metabolic activity (cleavage of dimethyl-thiazolyl-diphenyltetrazo-lium bromide) (MTT) and DNA synthesis (incorporation of 5-bromo-2 -deoxyur-idine [BrdU]) assay on L-929 mouse fibroblast cells. It was found that the toxicity of chitosan-TBA was concentration-dependent [63]. Another thiolated chitosan investigated for cytotoxicity was chitosan-MBA, which was found to be nontoxic because Caco-2 cell viability was around 100% after 4 and 24 h of incubation [25]. 

Another method is the red blood cell (lysis) test (RBC test) based on the (cytotoxic) potential of a chemical substance to disrupt cell membranes. The RBC test contributes to the reduction of animal numbers used for eye irritation testing. Access to mammalian erythrocytes is easy (e.g. slaughterhouse material). The RBC test is used in industry as part of an in vitro test battery, which is routinely used as a pre-screen (Brantom et al, 1997 Harbell et al, 1997). Membrane damage is assessed by photometrically measuring the leakage of haemoglobin from freshly isolated red blood cells incubated with test materials under standard conditions (Pape and Hoppe, 1991 Lewis et al, 1994). 

Red blood cell lysis, photosensitized by glipizide 9, and the products of its aerobic photolysis were demonstrated. The photohemolysis rate was lower for 9 than for its photoproducts. Inhibition of this... 

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