Quantitative study of starving platelets in a minimal medium: maintenance by acetate or plasma but not by glucose.
The requirement of donor platelets for fuels, plasma and calcium were studied using platelets washed, filtered to remove leucocytes and resuspended in a new glucose-free minimal platelet storage medium with low citrate (3 mmol/l), low buffer capacity and no calcium. This is the first study of platelets stored without plasma, glucose or calcium and it was shown that platelets continued to aggregate with collagen plus adrenaline for 48 h and showed only a 50% fall in 'swirl index', an objective morphology score, after 3 days, showing that by these criteria human platelets do not require glucose. Sodium acetate extended the storage time by between 2 and 4 days, depending on the index parameter. This is the first evidence showing that failure of platelets in these conditions is at least partly due to exhaustion of fuel, and the first evidence that acetate prolongs in vitro survival. As little as 10% low-glucose plasma extended the storage time, but it was no better than acetate. New observations using this system included a very rapid fall in pH during resuspension of the washed platelet pellet, a rising pH in the absence of added fuel and an increased pH with added acetate.
Whisson
M E
ME
Red Cross Blood Transfusion Service, Perth, Australia.
Nakhoul
A
A
Howman
P
P
Niu
X
X
Guppy
M
M
D020155:3-Hydroxybutyric Acid; D000085:Acetates / Q000494:pharmacology*; D019342:Acetic Acid; D001792:Blood Platelets / Q000187:drug effects* / Q000378:metabolism; D001793:Blood Preservation; D002122:Calcium Chloride / Q000378:metabolism / Q000494:pharmacology; D002470:Cell Survival / Q000187:drug effects; D016895:Culture Media, Serum-Free / Q000494:pharmacology*; D004734:Energy Metabolism; D005690:Galactose / Q000494:pharmacology; D005947:Glucose / Q000494:pharmacology*; D006003:Glycogen / Q000032:analysis; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006885:Hydroxybutyrates / Q000494:pharmacology; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D010101:Oxygen Consumption; D010949:Plasma; D010974:Platelet Aggregation; D010976:Platelet Count
D000085:Acetates; D016895:Culture Media, Serum-Free; D006885:Hydroxybutyrates; D006003:Glycogen; D007770:L-Lactate Dehydrogenase; D005947:Glucose; D002122:Calcium Chloride; D019342:Acetic Acid; D020155:3-Hydroxybutyric Acid; D005690:Galactose