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 pH i 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 V max 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 pH i regulation system is largely preserved during freeze-drying and rehydration of PLTs.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 1029-1037 |
| Number of pages | 9 |
| Journal | Transfusion |
| Volume | 46 |
| Issue number | 6 |
| DOIs | |
| State | Published - Jun 2006 |
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ASJC Scopus subject areas
- Hematology
- Immunology
Cite this
Freeze-dried rehydrated human blood platelets regulate intracellular pH. / Tang, Minke; Wolkers, Willem F.; Crowe, John H.; Tablin, Fern.
In: Transfusion, Vol. 46, No. 6, 06.2006, p. 1029-1037.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Freeze-dried rehydrated human blood platelets regulate intracellular pH
AU - Tang, Minke
AU - Wolkers, Willem F.
AU - Crowe, John H.
AU - Tablin, Fern
PY - 2006/6
Y1 - 2006/6
N2 - 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 pH i 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 V max 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 pH i regulation system is largely preserved during freeze-drying and rehydration of PLTs.
AB - 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 pH i 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 V max 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 pH i regulation system is largely preserved during freeze-drying and rehydration of PLTs.
UR - http://www.scopus.com/inward/record.url?scp=33744463370&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33744463370&partnerID=8YFLogxK
U2 - 10.1111/j.1537-2995.2006.00838.x
DO - 10.1111/j.1537-2995.2006.00838.x
M3 - Article
C2 - 16734821
AN - SCOPUS:33744463370
VL - 46
SP - 1029
EP - 1037
JO - Transfusion
JF - Transfusion
SN - 0041-1132
IS - 6
ER -