Preparation of low-affinity red cells with dimethylsulfoxide-mediated inositol hexaphosphate incorporation: hemoglobin and ATP recovery using a continuous-flow method.
Incorporation of IHP into red cells decreases oxygen affinity as a result of the binding of this compound to the 2,3-DPG site of hemoglobin. This investigation describes a continuous-flow method which utilizes the osmotic pulse technique to transport IHP into RBC. Using this procedure, it is possible to obtain a significant increase in P50 while maintaining in vitro cellular integrity. For example, IHP incorporation sufficient to cause an increase in the P50 of 20 mm Hg may be achieved with recovery of approximately 75% of the hemoglobin and with maintenance of ATP levels compatible with good viability. The continuous-flow method allows uniform treatment of large, unit-size volumes of red cells with a relatively small quantity of reagents. The final cell product is macrocytic/hypochromic with an increased number of stomatocytes.
D000255:Adenosine Triphosphate / Q000097:blood*; D002469:Cell Separation / Q000379:methods*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D004909:Erythrocyte Indices; D004912:Erythrocytes / Q000166:cytology / Q000378:metabolism*; D006454:Hemoglobins / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D010100:Oxygen / Q000097:blood; D010313:Partial Pressure; D010833:Phytic Acid / Q000097:blood*; D011092:Polyethylene Glycols / Q000494:pharmacology
D006454:Hemoglobins; D011092:Polyethylene Glycols; D010833:Phytic Acid; D000255:Adenosine Triphosphate; D010100:Oxygen; D004121:Dimethyl Sulfoxide