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Clinical use of frozen platelets
680
ABSTRACTS,
19th ANNUAL
scaled-up, closed, and disposable system of 250-ml consisting of three interconnected plastic bags suggest that this system can be developed for effective use in platelet transfusion. 98. Clinical
Use of Frozen Platelets.
S. SUMIDA
(De-
partment of Surgery, National Fukuoka Central Hospital, Jonai 2-2, Fukuoka 810, Japan). Within an hour after blood collection, platelet concentrates were frozen with the same volume of 10% glycerol solution (10% glycerol, 1% mannitol, 1% glucose, and 0.6% NaCl). The slow cooling rate was controlled at Z”C/min until -65”C, followed by immersion into liquid nitrogen. Thawed deglycerolized platelet concentrates were infused intravenously after one wash with 100 ml of 0.9% saline solution. The effect of thawed deglycerolized platelets was evaluated by the aspirin bleeding tendency and 51Crlife span after the autologous transfusion of processed platelets into the original donors. The frozen platelets improved the prolonged bleeding tendency from aspirin. The in vivo life spans (T& were 4.32 f 0.54 day (n = 5) for nonfrozen fresh platelets, and 3.48 f 1.11 (n = 9) for frozen platelets. The in vivo recovery rates were 72.8 f 8% for nonfrozen fresh platelets and 65 2 11% for frozen platelets, respectively. Blood cultures were 100%negative. No side effect from the transfusion was seen. Fifty or more units of frozen platelets in the frozen state in liquid nitrogen stocker were transported without damage from Fukuoka to Tokyo, and used for clinical patients. Five-hundred and ninety-five units of thawed platelets were infused into 59 patients complaining of thrombocytopenia and bleeding (oozing): 19 idiopathic thrombocytopenia purpura, 11 acute myeloic leukemia, and others. The platelet counts increased in 45 patients (76.3%), and the bleeding time (oozing) was improved in 38 patients (64.4%). HLA matching tests were performed for nine patients; however, a clear-cut difference was not observed in contrast with those patients who received frozen platelets without the HLA matching test. 99. Osmotic
Limits
of Human
Platelets.
MEETING
concentration dependence. Changes in pH were negligible. The morphological scores, however, showed a considerable decrease when PRP was exposed to solutions of 150 or 790 mOsm. The decrease was mainly caused by a shii from discoid to spherical or dendritic forms. After exposure to these osmolalities, aggregation of platelets induced by calcium ionophore (A23187) was similar to control, but aggregation induced by ADP and collagen as well as the hypotonic stress reversal were greatly impaired. Such results demonstrate that the tolerance of platelets to hyper and hypotonic stresses is limited. This suggests that care must be exercised in the introduction and removal of cryoprotectants during the platelet freezing process and that platelets might be expected to be more susceptible to the hypertonic stresses of freezing than other cells with broader osmotic tolerance. (Supported in part by NIH Grants HL 27535 and RR05737.) 100. Response of Platelets to Osmotic Stress and the Introduction
H. T.
and Removal
MERYMAN,
of Cryoprotectants.
M. ST. J. DOUGLAS,
AND P.
(ARC Blood Services Laboratories, Bethesda, Maryland 20814). LAW
Exposure to hyperosmotic solutions causes impairment of function in human platelets. Using morphological scoring as an assay we have examined the effect of creating hypertonicity by the addition of impermeant solutes other than NaCl. These included KCl, Na,SO,, (CH,),NCl, mannitol, fructose, and sucrose. None resulted in a significant improvement in morphological score on return to isotonicity as compared to cells exposed to solutions made hyperosmotic with NaCl. Pharmacological and other agents were tested for their intluence on hypertonic stress injury. These included calcium, magnesium, EDTA, aspirin, indomethacin, PGE,, chlorpromazine, polyvinylpyrrolidone, polyethylene glycol, and 0.1 M Me,SO. With the exception of the first three agents, all were modestly but not dramatically beneficial, improving scores after return to isotonicity by 5 to 50%. (Supported in part by NIH Grants HL 27535and RR05737.)
P. LAW,
M. ST. J. DOUGLAS, AND H. T. MERYMAN (ARC Blood Services Laboratories, Bethesda, Maryland 20814). Platelet-rich plasma (PRP) was prepared from single units of whole blood. PRP was loaded in dialysis tubing and exposed to various osmolalities of phosphatebuffered NaCl-dextrose solution at room temperature. Numerical recovery, morphological score, aggregation responses, and hypotonic stress reversal reaction were assayed as a function of osmolality after the PRP was returned to isotonicity. The decrease in numerical recovery was small (- 12%) and showed no
101. Treatment of Blast Crisis in Chronic Myelocytic Leukemia (CML) with Autologous Peripheral Cryopreserved Hemopoietic Stem Cells and Lymphocytes. R. SEELIS, M. W. SCHEIWE, U. ESSERS, Zs. PUSZTAI-MARKOS, AND K. H. STURNER (Medizinische Fakultat und Helm-
holtz-Institut fur Biomedizinische Technik, Goethestrasse 27/29, D-5100 Aachen, West Germany). In a clinical study cryopreserved autologous peripheral blood mononuclear cells (MNC) and hemopoietic stemcells (CFU-c) are used for improve-
ABSTRACTS,
19th ANNUAL
scaled-up, closed, and disposable system of 250-ml consisting of three interconnected plastic bags suggest that this system can be developed for effective use in platelet transfusion. 98. Clinical
Use of Frozen Platelets.
S. SUMIDA
(De-
partment of Surgery, National Fukuoka Central Hospital, Jonai 2-2, Fukuoka 810, Japan). Within an hour after blood collection, platelet concentrates were frozen with the same volume of 10% glycerol solution (10% glycerol, 1% mannitol, 1% glucose, and 0.6% NaCl). The slow cooling rate was controlled at Z”C/min until -65”C, followed by immersion into liquid nitrogen. Thawed deglycerolized platelet concentrates were infused intravenously after one wash with 100 ml of 0.9% saline solution. The effect of thawed deglycerolized platelets was evaluated by the aspirin bleeding tendency and 51Crlife span after the autologous transfusion of processed platelets into the original donors. The frozen platelets improved the prolonged bleeding tendency from aspirin. The in vivo life spans (T& were 4.32 f 0.54 day (n = 5) for nonfrozen fresh platelets, and 3.48 f 1.11 (n = 9) for frozen platelets. The in vivo recovery rates were 72.8 f 8% for nonfrozen fresh platelets and 65 2 11% for frozen platelets, respectively. Blood cultures were 100%negative. No side effect from the transfusion was seen. Fifty or more units of frozen platelets in the frozen state in liquid nitrogen stocker were transported without damage from Fukuoka to Tokyo, and used for clinical patients. Five-hundred and ninety-five units of thawed platelets were infused into 59 patients complaining of thrombocytopenia and bleeding (oozing): 19 idiopathic thrombocytopenia purpura, 11 acute myeloic leukemia, and others. The platelet counts increased in 45 patients (76.3%), and the bleeding time (oozing) was improved in 38 patients (64.4%). HLA matching tests were performed for nine patients; however, a clear-cut difference was not observed in contrast with those patients who received frozen platelets without the HLA matching test. 99. Osmotic
Limits
of Human
Platelets.
MEETING
concentration dependence. Changes in pH were negligible. The morphological scores, however, showed a considerable decrease when PRP was exposed to solutions of 150 or 790 mOsm. The decrease was mainly caused by a shii from discoid to spherical or dendritic forms. After exposure to these osmolalities, aggregation of platelets induced by calcium ionophore (A23187) was similar to control, but aggregation induced by ADP and collagen as well as the hypotonic stress reversal were greatly impaired. Such results demonstrate that the tolerance of platelets to hyper and hypotonic stresses is limited. This suggests that care must be exercised in the introduction and removal of cryoprotectants during the platelet freezing process and that platelets might be expected to be more susceptible to the hypertonic stresses of freezing than other cells with broader osmotic tolerance. (Supported in part by NIH Grants HL 27535 and RR05737.) 100. Response of Platelets to Osmotic Stress and the Introduction
H. T.
and Removal
MERYMAN,
of Cryoprotectants.
M. ST. J. DOUGLAS,
AND P.
(ARC Blood Services Laboratories, Bethesda, Maryland 20814). LAW
Exposure to hyperosmotic solutions causes impairment of function in human platelets. Using morphological scoring as an assay we have examined the effect of creating hypertonicity by the addition of impermeant solutes other than NaCl. These included KCl, Na,SO,, (CH,),NCl, mannitol, fructose, and sucrose. None resulted in a significant improvement in morphological score on return to isotonicity as compared to cells exposed to solutions made hyperosmotic with NaCl. Pharmacological and other agents were tested for their intluence on hypertonic stress injury. These included calcium, magnesium, EDTA, aspirin, indomethacin, PGE,, chlorpromazine, polyvinylpyrrolidone, polyethylene glycol, and 0.1 M Me,SO. With the exception of the first three agents, all were modestly but not dramatically beneficial, improving scores after return to isotonicity by 5 to 50%. (Supported in part by NIH Grants HL 27535and RR05737.)
P. LAW,
M. ST. J. DOUGLAS, AND H. T. MERYMAN (ARC Blood Services Laboratories, Bethesda, Maryland 20814). Platelet-rich plasma (PRP) was prepared from single units of whole blood. PRP was loaded in dialysis tubing and exposed to various osmolalities of phosphatebuffered NaCl-dextrose solution at room temperature. Numerical recovery, morphological score, aggregation responses, and hypotonic stress reversal reaction were assayed as a function of osmolality after the PRP was returned to isotonicity. The decrease in numerical recovery was small (- 12%) and showed no
101. Treatment of Blast Crisis in Chronic Myelocytic Leukemia (CML) with Autologous Peripheral Cryopreserved Hemopoietic Stem Cells and Lymphocytes. R. SEELIS, M. W. SCHEIWE, U. ESSERS, Zs. PUSZTAI-MARKOS, AND K. H. STURNER (Medizinische Fakultat und Helm-
holtz-Institut fur Biomedizinische Technik, Goethestrasse 27/29, D-5100 Aachen, West Germany). In a clinical study cryopreserved autologous peripheral blood mononuclear cells (MNC) and hemopoietic stemcells (CFU-c) are used for improve-