Preservation of Morphological Integrity and Clot Retraction Activity of Human Platelets After Freezing

1964 ◽  
Vol 11 (01) ◽  
pp. 222-229 ◽  
Author(s):  
Isaac Djerassi ◽  
Albert Roy ◽  
Jorge Alvarado ◽  
Keyword(s):  
Dimethyl Sulfoxide ◽  
Liquid Nitrogen ◽  
Human Platelets ◽  
Slow Freezing ◽  
Critical Conditions ◽  
Plasma Medium ◽  
Clot Retraction ◽  
Controlled Freezing ◽  
Direct Immersion

SummaryHuman platelets frozen at −195° C (liquid nitrogen) retain their morphological integrity and ability to promote clot retraction when 5% dimethyl-sulfoxide and 5% dextrose are added to the suspending plasma medium. Slow freezing was more effective than direct immersion in the liquid nitrogen. Although similar results may be achieved with dimethylsulfoxide alone with rigidly controlled freezing rates, the addition of sugars may permit freezing under less critical conditions.Dimethylsulfoxyd und 5% Dextrose dem Plasmamilieu hinzugefügt werden. Das langsame Einfrieren ist effektiver als das direkte Eintauchen in flüssigen Stickstoff. Obschon ähnliche Resultate mit Dimethylsulfoxyd allein unter exakter Kontrolle der Einfrierungsgeschwindig-keit erreicht werden können, erlaubt die Zugabe von Dextrose ein Einfrieren unter weniger kritischen Bedingungen.

scholarly journals CRYOPRESERVATION OF DORMANT GRAPE (Vitis sp.) BUDS.

HortScience ◽  
1990 ◽  
Vol 25 (9) ◽  
pp. 1090e-1090 ◽  
Author(s):  
Virgil Esensee ◽  
Cecil Stushnoff ◽  
Philip L. Forsline
Keyword(s):  
Liquid Nitrogen ◽  
Electrolyte Leakage ◽  
Slow Freezing ◽  
First Report ◽  
Vitis Sp ◽  
Direct Immersion ◽  
Backup Storage

There is need for backup storage of clonally propagated plant cultivars of numerous taxa. Initial tests, using a protocol developed for dormant apple buds that includes desiccation and slow freezing prior to immersion in liquid nitrogen (-196 C), was not effective with `Valiant' grape. Accordingly, replicates of V. vinifera `Riesling', V. riparia, `Valiant' and a V. amurensis × riparia cross were also tested for survival at –196 C, following desiccation to 25% & 18% water (fwb) and direct immersion into liquid nitrogan. Visual and electrolyte leakage ratings following nine days of dehydration in moist peat were used to assess viability. Direct immersion of desiccated samples resulted in survival for some buds of `Valiant' and a V. amurensis × riparia cross. V. riparia showed some survival when field hydrated and at 25% water, while all buds desiccated to 18% survived. `Riesling' did not survive desiccation, and was killed by all -196 C treatments. The apple protocol was partially effective, in combination with desiccation to 18% in `Valiant' and V. riparia. This is the first report of grape bud survival in liquid nitrogen and more detailed studies are planned.

Release, Aggregation and Lysis of Human Platelets by Antilymphocyte Globulin and Antiplatelet Serum

1976 ◽  
Vol 36 (02) ◽  
pp. 411-423 ◽  
Author(s):  
Nicholas Lekas ◽  
J. C Rosenberg
Keyword(s):  
Platelet Aggregation ◽  
Gamma Globulin ◽  
Human Platelets ◽  
Plasma Medium ◽  
Release Reaction ◽  
Clinical Events ◽  
Thrombotic Complications ◽  
Platelet Release ◽  
Free Media

SummaryHuman platelets labeled with 51Cr were used to determine the contribution made by platelet lysis to the platelet release reaction and platelet aggregation induced by rabbit antihuman platelet serum (APS) and equine antihuman thymocyte globulin (ATG). Platelets were tested in both plasma (PRP) and non-plasma containing media. Antibodies directed against platelets, either as APS or ATG, induced significant amounts of platelet release and aggregation, as well as some degree of lysis, in the absence of complement. The presence of complement increased platelet lysis and aggregation, but not the release reaction. Non-immune horse gamma globulin produced different responses depending upon whether platelets were investigated in PRP or non-plasma containing media. Aggregation was seen in the latter but not the former. These differences can be explained by the presence of plasma components which prevent non-specific immune complexes from causing platelet aggregation. Since platelets in vivo are always in a plasma medium, one must be wary of utilizing data from platelet studies in synthetic plasma-free media as the basis of explaining clinical events. These observations demonstrate at least two, and possibly three, different mechanisms whereby ATG could activate platelets causing thrombotic complications and thrombocytopenia, i.e., via 1) specific and, 2) non-specific non-lytic pathways and 3) a lytic pathway.

Fetal Serum in Combination with 5% Dimethyl Sulfoxide Efficiently Protects the Human Gut Microbiota during Cryopreservation in Liquid Nitrogen

BIOPHYSICS ◽  
2021 ◽  
Vol 66 (4) ◽  
pp. 657-664
Author(s):  
L. V. Zalomova ◽  
D. A. Reshetnikov ◽  
S. V. Ugraitskaya ◽  
L. M. Mezhevikina ◽  
A. V. Zagainova ◽  
...  
Keyword(s):  
Gut Microbiota ◽  
Dimethyl Sulfoxide ◽  
Liquid Nitrogen ◽  
Human Gut ◽  
Human Gut Microbiota ◽  
Fetal Serum

Abstract 13598: The G-protein BetaGamma Subunits Regulate Platelet Function

Circulation ◽  
2020 ◽  
Vol 142 (Suppl_3) ◽  
Author(s):  
Ahmed Alarabi ◽  
Zubair Karim ◽  
Victoria Hinojos ◽  
Patricia A Lozano ◽  
Keziah Hernandez ◽  
...  
Keyword(s):  
G Protein ◽  
Platelet Function ◽  
Thrombus Formation ◽  
Human Platelets ◽  
Inhibitory Effects ◽  
Clot Retraction ◽  
Betagamma Subunits

Platelet activation involves tightly regulated processes to ensure a proper hemostasis response, but when unbalanced, can lead to pathological consequences such as thrombus formation. G-protein coupled receptors (GPCRs) regulate platelet function by interacting with and mediating the response to various physiological agonists. To this end, an essential mediator of GPCR signaling is the G protein Gαβγ heterotrimers, in which the βγ subunits are central players in downstream signaling pathways. While much is known regarding the role of the Gα subunit in platelet function, that of the βγ remains poorly understood. Therefore, we investigated the role of Gβγ subunits in platelet function using a Gβγ (small molecule) inhibitor, namely gallein. We observed that gallein inhibits platelet aggregation and secretion in response to agonist stimulation, in both mouse and human platelets. Furthermore, gallein also exerted inhibitory effects on integrin αIIbβ3 activation and clot retraction. Finally, gallein’s inhibitory effects manifested in vivo , as documented by its ability to modulate physiological hemostasis and delay thrombus formation. Taken together, our findings demonstrate, for the first time, that Gβγ directly regulates GPCR-dependent platelet function, in vitro and in vivo . Moreover, these data highlight Gβγ as a novel therapeutic target for managing thrombotic disorders.

scholarly journals Dasatinib Inhibits Procoagulant and Clot Retracting Activities of Human Platelets

2019 ◽  
Vol 20 (21) ◽  
pp. 5430
Author(s):  
Ildikó Beke Debreceni ◽  
Gabriella Mezei ◽  
Péter Batár ◽  
Árpád Illés ◽  
János Kappelmayer
Keyword(s):  
Chronic Myeloid Leukemia ◽  
Western Blot ◽  
Myeloid Leukemia ◽  
Thrombin Generation ◽  
Platelet Adhesion ◽  
Kinase Inhibitors ◽  
Human Platelets ◽  
Activation Loop ◽  
Clot Retraction ◽  
Lines Of Evidence

Tyrosine kinase inhibitors (TKI) such as the BCR-ABL inhibitor dasatinib and nilotinib are highly effective therapies for chronic myeloid leukemia (CML). However, several lines of evidence suggest that dasatinib can induce bleeding which may be due to impaired collagen-induced platelet adhesion, aggregation, and secretion. Sarcoma family kinases (SFK) play central role in the GPVI-induced signaling pathway. We aimed to investigate whether and how dasatinib can modulate SFK-mediated platelet procoagulant activity in a purified system and in dasatinib/nilotinib treated CML patients. In platelet rich plasmas of healthy volunteers, dasatinib dose-dependently reduced convulxin-induced phosphatidylserine exposure and attenuated thrombin formation. Similarly to these changes, integrin activation and clot retraction were also significantly inhibited by 100 nM dasatinib. Platelets isolated from dasatinib treated patients showed a significantly lower phosphatidylserine expression upon convulxin activation compared to premedication levels. In these samples, thrombin generation was significantly slower, and the quantity of formed thrombin was less compared to the trough sample. Western blot analyses showed decreased phosphorylation levels of the C-terminal tail and the activation loop of SFKs upon dasatinib administration. Taken together, these results suggest that dasatinib inhibits the formation of procoagulant platelets via the GPVI receptor by inhibiting phosphorylation of SFKs.

Cryopreservation of human platelets with dimethyl sulfoxide: changes in biochemistry and cell function

Transfusion ◽  
1995 ◽  
Vol 35 (11) ◽  
pp. 921-924 ◽  
Author(s):  
M Bock ◽  
M Schleuning ◽  
MU Heim ◽  
W Mempel
Keyword(s):  
Dimethyl Sulfoxide ◽  
Cell Function ◽  
Human Platelets

scholarly journals Survival and recovery of human platelets stored for five days in a non- plasma medium

Blood ◽  
1986 ◽  
Vol 67 (3) ◽  
pp. 672-675 ◽  
Author(s):  
GA Adams ◽  
SD Swenson ◽  
G Rock
Keyword(s):  
Platelet Aggregation ◽  
Human Blood ◽  
Blood Platelets ◽  
Human Platelets ◽  
Plasma Medium ◽  
Release Reaction ◽  
In Vivo Recovery ◽  
Human Blood Platelets

Abstract Human blood platelets were stored for five days as concentrates in 60 mL of: (a) plasma; (b) non-plasma medium with anticoagulant; and (c) non-plasma medium without anticoagulant. All preparations were equally functional when tested for platelet aggregation and release reaction in response to single agonist or synergistic pairs of agonists in vitro. Platelets stored in non-plasma medium with anti-coagulant had lower kallikrein, fibrino(gen)peptide A, lactate, and beta-thromboglobulin than did plasma controls after five days. In vivo recovery and survival of platelets stored in non-plasma medium with anticoagulant were 51.2% +/- 4.3% and 8.7 +/- 0.3 days, respectively, which were not statistically different from plasma controls of 39.2% +/- 4.9% and 7.2 +/- 0.8 days, respectively. It is concluded that platelets can be stored for five days in a non-plasma medium and still have good in vivo recoveries and survivals.

scholarly journals Spermatic abnormalities of piracanjuba Brycon orbignyanus (Valenciennes, 1849) after cryopreservation

2011 ◽  
Vol 71 (3) ◽  
pp. 693-699 ◽  
Author(s):  
JM. Galo ◽  
DP. Streit-Junior ◽  
RN. Sirol ◽  
RP. Ribeiro ◽  
M. Digmayer ◽  
...  
Keyword(s):  
Dimethyl Sulfoxide ◽  
Liquid Nitrogen ◽  
Distilled Water ◽  
Reproductive Characteristics ◽  
Fish Sperm ◽  
Before And After ◽  
Cryoprotectant Solution ◽  
Fresh Semen

The objective of this research was to verify the presence of spermatic abnormalities on semen of Brycon orbignyanus after cryopreservation. Semen was collected from ten four-year-old males who presented secondary reproductive characteristics for migrating fish. Sperm was evaluated for motility, vigor and spermatic morphology before and after cryopreservation. A cryoprotectant solution was made of 20 mL of yolk egg, 5.0 g of glucose and dimethyl sulfoxide diluted in distilled water (10 mL: 90 mL). The diluted semen (1:3, semen:solution) was submitted to nitrogen steam for 24 hours and then to liquid nitrogen (-196 ºC) for 60 days. Cryopreservation decreased the percentage of normal spermatozoa from 62.20% to 54.60%. Consequently, the percentage of spermatozoa with secondary abnormalities increased from 8.50% to 15.00%. However, there was no difference in primary abnormalities. Both spermatic motility and vigor were decreased in cryopreserved semen compared with fresh semen. In conclusion, cryopreservation of semen of B. orbignyanus increased the percentage of secondary abnormalities and decreased the spermatic motility and vigor.

AMPK α2 subunit is involved in platelet signaling, clot retraction, and thrombus stability

Blood ◽  
2010 ◽  
Vol 116 (12) ◽  
pp. 2134-2140 ◽  
Author(s):  
Voahanginirina Randriamboavonjy ◽  
Johann Isaak ◽  
Timo Frömel ◽  
Benoit Viollet ◽  
Beate Fisslthaler ◽  
...  
Keyword(s):  
Kinase Inhibitors ◽  
Adenosine Monophosphate ◽  
Human Platelets ◽  
Whole Body ◽  
Initial Increase ◽  
Clot Retraction ◽  
Compound C ◽  
Platelet Signaling ◽  
Upstream Kinase

Abstract The adenosine monophosphate (AMP)–activated protein kinase (AMPK) is a regulator of energy balance at the cellular and whole-body levels, but little is known about the role of AMPK in platelet activation. We report that both the α1 and α2 AMPK isoforms are expressed by human and murine platelets and that thrombin elicits the phosphorylation of AMPKα as well as the upstream kinase, liver kinase B1 (LKB1). In human platelets, the kinase inhibitors iodotubercidin and compound C significantly inhibited thrombin-induced platelet aggregation and clot retraction without affecting the initial increase in [Ca2+]i. Clot retraction was also impaired in platelets from AMPKα2−/− mice but not from wild-type littermates or AMPKα1−/− mice. Moreover, rebleeding was more frequent in AMPKα2−/− mice, and the FeCl3-induced thrombi formed in AMPKα2−/− mice were unstable. Mechanistically, AMPKα2 was found to phosphorylate in vitro the Src-family kinase, Fyn, and isoform deletion resulted in the attenuated threonine phosphorylation of Fyn as well as the subsequent tyrosine phosphorylation of its substrate, β3 integrin. These data indicate that AMPKα2—by affecting Fyn phosphorylation and activity—plays a key role in platelet αIIbβ3 integrin signaling, leading to clot retraction and thrombus stability.

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