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- Title
Freeze-dried rehydrated human blood platelets regulate intracellular pH.
- Authors
Tang, Minke; Wolkers, Willem F.; Crowe, John H.; Tablin, Fern
- Abstract
BACKGROUND: Long-term storage of platelets (PLTs) in the dry state would greatly improve options for PLT storage. Whether trehalose-loaded freeze-dried and rehydrated PLTs could regulate intracellular pH (pHi) was evaluated. STUDY DESIGN AND METHODS: Previously it was shown that human PLTs can be successfully preserved by freeze-drying with trehalose. Trehalose-loaded freeze-dried rehydrated PLTs and fresh control PLTs were labeled with the pH dye BCECF-AM. pHi was measured in resting cells, cells acidified with nigericin, and cells treated with thrombin. The sodium-proton pump was blocked by treatment with 5-( N-methyl- N-isobutyl)amiloride (MIA). RESULTS: The pHi of rehydrated PLTs is the same as that of fresh control PLTs, 7.27 ± 0.03 (SD; n = 5) and 7.27 ± 0.02 (n = 5), respectively. Nigericin treatment of cells showed that the recovery in pHi was Na+-dependent and followed Michaelis-Menten kinetics. The Vmax values (ΔpH/9 sec) were 0.21 ± 0.039 (n = 3) and 0.22 ± 0.025 (n = 3) for rehydrated and control PLTs, respectively. The exchange constants were 17.7 ± 2.3 mmol per L (n = 3) and 17.0 ± 1.9 mmol per L (n = 3) for rehydrated and control PLTs, respectively. Treatment of cells with MIA showed that NHE1 remained sensitive to the inhibitor after freeze-drying and rehydration. CONCLUSION: The results show that the pHi regulation system is largely preserved during freeze-drying and rehydration of PLTs.
- Subjects
BLOOD platelets; FREEZE-drying; THROMBIN; CYTOLOGY; PROTON pump inhibitors; AMILORIDE
- Publication
Transfusion, 2006, Vol 46, Issue 6, p1029
- ISSN
0041-1132
- Publication type
Article
- DOI
10.1111/j.1537-2995.2006.00838.x