Improved stability of 2,3-bisphosphoglycerate during storage of hexokinase-overloaded erythrocytes.
Human red blood cells were overloaded with homogeneous human hexokinase using a procedure of encapsulation based on hypotonic hemolysis and isotonic resealing and reannealing to achieve a final activity that was 15 times higher than that in control cells. Storage for 5 weeks at 4 degrees C of hexokinase-overloaded erythrocytes shows that these cells undergo small K+ leakage and mean cell volume increase compared with control cells. Furthermore, after these 5 weeks of storage the 2,3-bisphosphoglycerate content was normal while the ATP concentration was slightly reduced. These results and other properties suggest that encapsulation of key glycolytic enzymes in erythrocytes can provide a new way to maintain in vitro functionally active red blood cells for at least 5 weeks.
Magnani
M
M
Instituto di Chimica Biologica, Università degli Studi, Urbino, Italy.
Rossi
L
L
Bianchi
M
M
Serafini
G
G
Zocchi
E
E
Laguerre
M
M
Ropars
C
C
D019794:2,3-Diphosphoglycerate; D000255:Adenosine Triphosphate / Q000097:blood; D004163:Diphosphoglyceric Acids / Q000097:blood*; D004912:Erythrocytes / Q000201:enzymology*; D006593:Hexokinase / Q000097:blood*; D006801:Humans; D066298:In Vitro Techniques
D004163:Diphosphoglyceric Acids; D019794:2,3-Diphosphoglycerate; D000255:Adenosine Triphosphate; D006593:Hexokinase