Preparation Of Cryoprecipitate Research Articles

A comparative evaluation of the quality of cryoprecipitate prepared from 350ml versus 450ml of whole blood and different methods of thawing of plasma: A prospective observational study.

Cryoprecipitate is used in conditions like hypofibrinogenemia, massive transfusion with bleeding, and factor XIII deficiency. The current guidelines support the preparation of cryoprecipitate from 450ml whole blood. But 350ml of whole blood collection is expected from low body weight (<55kg) donors. However, no standardized criteria exist for preparing cryoprecipitate from 350ml of whole blood. This study compared the fibrinogen and factor VIII levels in cryoprecipitate units prepared from 350ml versus 450ml whole blood collection. The study also compared the fibrinogen and factor VIII levels prepared by circulating water bath versus blood bank refrigerator (BBR) thawing method. A total of 128 blood bags were equally divided into groups A and B for 450 and 350ml whole blood collection further subdivided into subgroups based on thawing methods. The fibrinogen and factor VIII yield were analyzed in the cryoprecipitates prepared from both groups. The factor VIII levels were significantly higher in cryoprecipitate made from 450ml whole blood collection (P=0.02). The BBR method of plasma thawing resulted in better fibrinogen recovery than the cryo bath method. Whereas vice versa in the case of factor VIII recovery. A weak but significant positive correlation was noted in factor VIII levels with the plasma volume. Over 75% of the cryoprecipitates prepared from 350ml whole blood passed the fibrinogen and factor VIII quality control criteria. So, 350ml whole blood collection from low body weight (<55kg) donors could be utilized to prepare cryoprcipitates. However, future clinical studies should focus on the cryoprecipitate's clinical efficacy prepared from 350ml of whole blood.

Preparation of Cryoprecipitate and Cryo-depleted Plasma for Proteomic Research Analysis.

When frozen plasma is slowly thawed in cold conditions (1-6°C), high-molecular-weight plasma proteins precipitate forming a concentrate known as cryoprecipitate. The concentrate is enriched with several important coagulation proteins, including fibrinogen, antihemophilic factor (factor VIII), von Willebrand factor, fibrin stabilizing factor (factor XIII), fibronectin, and small amounts of other plasma proteins. In current medical practice, clinical-grade preparations of cryoprecipitate are used mostly to correct fibrinogen deficiency caused by acute blood loss or due to functional abnormalities of the fibrinogen protein. In the past, cryoprecipitate was used to treat von Willebrand disease and hemophilia A (factor VIII deficiency), but the availability of more highly purified coagulation factor concentrates or recombinant protein preparations has superseded the use of cryoprecipitate for these coagulopathies. Cryo-depleted plasma (also called cryosupernatant) is the plasma supernatant remaining following removal of the cryoprecipitate from frozen-thawed plasma and contains all the remaining soluble plasma proteins. This protocol describes the research-scale preparation of cryoprecipitate and cryo-depleted plasma suitable for proteomic studies and is based on the procedures used to prepare clinical-grade cryoprecipitate.

Preparation and Storage of Cryoprecipitate Derived from Amotosalen and UVA-Treated Apheresis Plasma and Assessment of In Vitro Quality Parameters.

Cryoprecipitate is a plasma-derived blood product, enriched for fibrinogen, factor VIII, factor XIII, and von Willebrand factor. Due to infectious risk, the use of cryoprecipitate in Central Europe diminished over the last decades. However, after the introduction of various pathogen-reduction technologies for plasma, cryoprecipitate production in blood centers is a feasible alternative to pharmaceutical fibrinogen concentrate with a high safety profile. In our study, we evaluated the feasibility of the production of twenty-four cryoprecipitate units from pools of two units of apheresis plasma pathogen reduced using amotosalen and ultraviolet light A (UVA) (INTERCEPT® Blood System). The aim was to assess the compliance of the pathogen-reduced cryoprecipitate with the European Directorate for the Quality of Medicines (EDQM) guidelines and the stability of coagulation factors after frozen (≤−25 °C) storage and five-day liquid storage at ambient temperature post-thawing. All pathogen-reduced cryoprecipitate units fulfilled the European requirements for fibrinogen, factor VIII and von Willebrand factor content post-preparation. After five days of liquid storage, content of these factors exceeded the minimum values in the European requirements and the content of other factors was sufficient. Our method of production of cryoprecipitate using pathogen-reduced apheresis plasma in a jumbo bag is feasible and efficient.

Effects and issues of in-hospital preparation of cryoprecipitates

Effects and issues of in-hospital preparation of cryoprecipitates

Clinical guidelines for cryosupernatant transfusions

Introduction. Cryosupernatant is blood component. Cryosupernatant is the supernatant plasma removed during the preparation of cryoprecipitate. Aim. To provide information on the composition and methods of production, storage, transportation and clinical use of Cryosupernatant. General fi ndings. In comparison with fresh frozen plasma (FFP) and cryoprecipitate, Cryosupernatant plasma is depleted in factor VIII, fi brinogen factor von Willebrand (VWF). Cryosupernatant is defi cient in high molecular weight multimers of VWF, but contains VWF metalloproteinase. The concentrations of factor V, antithrombin III, albumin and immunoglobulins are the same as in FFP and cryoprecipitate. The indications for Cryosupernatant transfusions are massive blood loss in patients with factor VIII inhibitor, plasma exchange in patients with thrombotic thrombocytopenic purpura. For children the doses of Cryosupernatant should be 10-15 mL/kg.

The Impact of Thromboelastography on Blood Transfusion Policy in Adult Cardiac Surgery-A Retrospective Observational Study from Eastern India.

Thisstudy was conducted todetermine the impact of thromboelastography (TEG) on blood transfusion policy regarding utilization and preparation of cryoprecipitate in adult cardiac surgery. The differences in total transfusion requirement, length of postoperative ICU stay and 24h mortality were also studied after introduction of TEG in transfusion protocol. It was a retrospective, single-center, observational study conducted in adult patients underwent cardiac surgery from April 2008 to March 2016. Two thousand patients underwent surgery when TEG was used compared with 1000 control patients beforeavailability of TEG. Significantly more patients in the TEG group versus the control group received cryoprecipitate (41 vs. 7%; p < 0.05), while fewer received a transfusion (60 vs. 87.5%; p < 0.05). Significant increase in cryoprecipitate preparation was observed after introduction of TEG. Patients underwent surgery in TEG group showed substantial reduction in administration of PRBC (2.1 vs. 3.5 U; p < 0.05); FFP (2.4 vs. 3.8 U; p < 0.05) and platelets (1.1 vs. 2.7 U; p < 0.05) compared to control group without compromising the length of ICU stay or postoperative mortality. A TEG-guided approach in adult patients undergoing cardiac surgery may increase the use of cryoprecipitate, while decreasing the overall requirement of blood transfusion.

A global initiative on local preparation of virus inactivated cryoprecipitate in developing countries

In developing countries, treatment of patients with bleeding disorders is inadequate or absent and the dramatic situation has not changed significantly during the past 20 to 30 years despite several initiatives set up by different national and international organizations. Past experience has shown that industrially manufactured clotting factor concentrates cannot solve these problems in developing countries, if they are used as the sole means in the treatment of patients with bleeding disorders. Recently, significant developments have occurred which have the potential to change the disastrous situation in therapy delivery to “hemophiliacs” in developing countries: blood systems (with blood services and blood centers) in many resource limited countries have tangibly improved and novel technologies for viral inactivation of blood components have been developed and are marketed [amotosalen, riboflavin, solvent-detergent (SD), etc.]. In a few developing countries, local preparation of viral inactivated cryoprecipitate (Cryo-VI) has been introduced and Cryo-VI has proven to be a safe, effective therapeutic for patients with bleeding disorders, at an affordable cost. But the use of VI-technology to render cryoprecipitate safe remains very limited; therefore, major coordinated actions are needed to promote Cryo-VI in developing countries and to facilitate implementation of local preparation of safe cryoprecipitate. Therefore, a Global Initiative on “local preparation of viral inactivated cryoprecipitate in developing countries” (“Local Cryo-VI in l-HDI”) has been launched to improve treatment of patients with bleeding disorders: it is based on collaboration with international partnering organizations (like WHO, ISBT and others) and national stakeholders (e.g., competent authorities, blood suppliers, patient associations). The Global Initiative consists of six core interventions, flanked by additional activities: revise and update existing standards and guidelines on “anti-hemophilic” treatment; give high priority in policies and strategies to “Local Cryo-VI in l-HDI”; organize strong advocacy; run a Pilot Project (PP) in several pilot sites; establish an expansion programme; support activities for implementation and sustainability of “Local Cryo-VI in l-HDI”. It is complementary to existing relief activities (e.g., product donations) and will result in additional supply of safe and effective haemostatic products to treat patients with bleeding disorders in developing countries. It will also contribute to significantly lower the incidence of inhibitors in previously untreated patients with hemophilia A.

Factors affecting the quality of cryoprecipitate.

BACKGROUND:Many variables affect the quality of cryoprecipitate (CRYO). We investigated the effect of freezing techniques and ABO blood groups on the quality of CRYO with respect to factor VIII: C and fibrinogen levels.MATERIALS AND METHODS:Ninety-six whole blood units each collected from in-house (Group I) and blood donation camps outside the hospital premises (Group II) were processed for CRYO preparation. Within each group, half the number of plasma units was frozen using blast freezer and another half using the conventional freezer. The CRYOs from blood groups A, B, and O were equally distributed, i.e. 32 within each of the Groups I and II. The fibrinogen and factor VIII: C levels in CRYO were analyzed using single-stage clotting assay.RESULTS:In Group I, the mean ± standard deviation percentage recovery of factor VIII levels in CRYO prepared using the conventional freezer and blast freezer were 58.5% ±16.2% and 66.7% ±16.4%, respectively, and in Group II, it was 55.3% ±17.6% and 70.4% ±13.4%, respectively. Recovery of factor VIII was higher in CRYO prepared using blast freezer than that of CRYO prepared using conventional freezer (P < 0.000). In Group II, CRYOs prepared using blast freezer had higher percent recovery of fibrinogen than that of Group I. In both the groups, the mean factor VIII levels in blood group A were higher than that of factor VIII levels in the blood group O CRYO.CONCLUSION:The factor VIII recovery in CRYO improves significantly with higher baseline factor VIII: C levels, blood group A donor, and rapid freezing using blast freezer. Rapid freezing also increases the fibrinogen yield.

A Protocol for the Preparation of Cryoprecipitate and Cryo-depleted Plasma for Proteomic Studies.

Cryoprecipitate is a concentrate of high-molecular-weight plasma proteins that precipitate when frozen plasma is slowly thawed at 1-6 °C. The concentrate contains factor VIII (antihemophilic factor), von Willebrand factor (vWF), fibrinogen, factor XIII, fibronectin, and small amounts of other plasma proteins. Clinical grade preparations of cryoprecipitate are mainly used to treat fibrinogen deficiency caused by acute bleeding or functional abnormalities of the fibrinogen protein. In the past, cryoprecipitate was used to treat von Willebrand disease and hemophilia A (factor VIII deficiency), but the availability of more highly purified coagulation factor concentrates or recombinant protein preparations has superseded the use of cryoprecipitate for these coagulopathies. Cryo-depleted plasma ("cryosupernatant") is the plasma supernatant remaining following removal of the cryoprecipitate from frozen-thawed plasma. It contains all the other plasma proteins and clotting factors present in plasma that remain soluble during cold-temperature thawing of the plasma. This protocol describes the clinical-scale preparation of cryoprecipitate and cryo-depleted plasma for proteomic studies.

Influencing Factors Control of Cryoprecipitate Preparation by Centrifugation

Objective To reduce the loss of coagulation factors by influencing factors control of cryoprecipitate preparation by centrifugation.Methods From July 2013 to June 2014,a total of 60 samples of cryoprecipitate prepared by centrifugation with strict control of influencing factors at Jingmen Red Cross Blood Center were selected into this study via simple random sampling method,as study group (n =60).From July 2012 to June 2013,a total of 54 samples of cryoprecipitate prepared by general centrifugation approach were selected into control group via simple random sampling method (n=54).The concentrations of factor Ⅷ (F Ⅷ) and fibronectin (Fib) in cryoprecipitate were analyzed.Comparison of average concentrations of FⅧ and Fib in cryoprecipitate between study group and control group were retrospectively analyzed.Results Average concentration of FⅧ and Fib in cryoprecipitate of study group and control group were both qualified.Average concentration of FⅧ in study group was (102.8±12.3) IU/bag,and average concentration of FⅧ in control group was (90.4 ± 4.2) IU/bag.Average concentration of F Ⅷ in study group was significantly higher than that in control group (t =8.04,P＜0.001).Average concentration of Fib in study group was (179.9 ± 6.7) mg/bag,and average concentration of Fib in control group was (170.2± 8.3) mg/bag.Average concentration of Fib in study group was also significantly higher than that in control group (t =7.72,P ＜0.01).Conclusion Strict control of influencing factors during cryoprecipitate preparation by centrifugation can reduce the loss of concentrations of FⅧ and Fib. Key words: Centrifugation; Cryoprecipitate coagulum; Factor Ⅷ; Fibronectin; Quality control

Preparation of cryoprecipitate from riboflavin and UV light-treated plasma

Background and objectivesThe Mirasol® pathogen reduction technology system for plasma is based on a riboflavin and UV light treatment process resulting in pathogen inactivation due to irreversible, photochemically induced damage of nucleic acids. This study was undertaken to evaluate the possibility of making pathogen reduced cryoprecipitate from riboflavin and UV light- treated plasma that meets the quality requirements specified by UK and European guidelines for untreated cryoprecipitate. Materials and methodsCryoprecipitate was made from riboflavin and UV light-treated plasma. Plasma units were thawed over a 20h period at 4°C, and variable centrifugation settings (from 654g for 2min to 5316g for 6min) were applied to identify the optimal centrifugation condition. Plasma proteins in cryoprecipitate units were characterized on a STA Compact, Diagnostica STAGO and Siemens BCS analyzer. ResultsNeither the centrifugation speed or time appeared to have an effect on the quality of the final cryoprecipitate product; however the initial solubilization of the cryoprecipitate product was found to be easier at the lower centrifugation setting (654g for 2min). Cryoprecipitate units prepared from Mirasol-treated plasma demonstrated protein levels that were less than levels in untreated products, but were on average 93IU/unit, 262mg/unit and 250IU/unit for FVIII, fibrinogen and von Willebrand ristocetin cofactor activity, respectively. ConclusionCryoprecipitate products prepared from Mirasol-treated plasma using a centrifugation method contain levels of fibrinogen, FVIII and von Willebrand ristocetin cofactor activity, that meet both the European and UK guidelines for untreated cryoprecipitate. Flexibility in centrifugation conditions should allow blood banks to use their established centrifugation settings to make cryoprecipitate from Mirasol-treated plasma.

Cees Th. Smit Sibinga: An Autobiography

Cees Smit Sibinga was born in 1939 in Makassar in the former Dutch East Indies and spent the first part of his life in a Japanese internment camp. This experience was extremely important for the development of his attitude in and philosophy of life, in which a positive fighter spirit predominates. He subsequently studied medicine, specializing in clinical hematology, and then did his PhD at the University of Groningen. In 1976, he became the Regional Blood Bank Director for the northern part of the Netherlands, which he shaped into a model blood bank. Since 1976, for 28 years, he annually organized an international symposium on various aspects of blood transfusion. Areas of his investigations included the development of a freeze-dried cryoprecipitate preparation used for hemophilia treatment, cryopreservation of stem cells and platelets, and quality management. His academic focus then turned to the teaching and training of transfusion personnel in resource-limited countries.

Cees Th. Smit Sibinga

Cees Smit Sibinga was born in 1939 in Makassar in the former Dutch East Indies and spent the first part of his life in a Japanese internment camp. This experience was extremely important for the development of his attitude in and philosophy of life, in which a positive fighter spirit predominates. He subsequently studied medicine, specializing in clinical hematology, and then did his PhD at the University of Groningen. In 1976, he became the Regional Blood Bank Director for the northern part of the Netherlands, which he shaped into a model blood bank. Since 1976, for 28 years, he annually organized an international symposium on various aspects of blood transfusion. Areas of his investigations included the development of a freeze-dried cryoprecipitate preparation used for hemophilia treatment, cryopreservation of stem cells and platelets, and quality management. His academic focus then turned to the teaching and training of transfusion personnel in resource-limited countries.

Preparation and viral inactivation of cryoprecipitate in blood banks in resource‐limited countries

Non‐virally inactivated cryoprecipitate prepared by blood banks is still used in many countries, especially in the developing world, for the treatment of bleeding disorders due to factor VIII (FVIII), von Willebrand factor (VWF), or fibrinogen deficiency. As a consequence, patients receiving cryoprecipitate are at high risk of being infected by plasma‐borne viruses, such as HIV, and hepatitis B virus (HBV) or C virus (HCV). It is therefore urgent to develop easy‐to‐use, affordable and efficient methods for the viral inactivation of cryoprecipitate under conditions that ensure good maintenance of its haemostatic properties. Preparation of cryoprecipitate from large‐pool solvent‐detergent (SD) plasma seems not economically or technically feasible in developing countries. Cryoprecipitate prepared from single‐donor plasma virally inactivated by methylene‐blue/illumination has significantly reduced content of factor VIII and fibrinogen. There are no data on the quality of cryoprecipitate produced from plasma virally inactivated by techniques in development such as S‐51 psoralen/UV irradiation or by riboflavin/UV irradiation, but loss of FVIII and fibrinogen is reported in S‐51/UV‐treated plasma. Dry‐heat treatments of smallpool cryoprecipitate or intermediate‐purity factor VIII have been developed for use by blood banks, but they are not much used and some have not stopped the risk of transmission of HCV. Recently, a mini‐pool process and a closed bag system to perform SD treatment of cryoprecipitate have been developed for use by blood banks. Recovery of factor VIII, VWF, and fibrinogen is over 95%. Viral validation studies revealed reduction factors of &gt; 4·17, &gt; 4·73, and &gt; 4·72 for HIV, BVDV, and PRV, respectively, within a few minutes of treatment. The SD bag system is in the process of CE‐marking and field testing should start in the near future.

Transfusion Protocol for Implantation and Explantation of Cardiac Devices in an Academic Medical Center

Few reports in the literature describe the transfusion support needed for implantation and explantation of cardiac assist devices. Because such devices are frequently used at this institution, we have established a protocol to support these patients. We reviewed blood bank records to document the usage of blood components in all patients on cardiac assist devices from 1989 to 2005 (n = 215). The data were separated into components given at the time of implantation and explantation of the device. Implantation data on 214 patients showed the mean use as follows: red blood cells (RBC), 5.7 U; platelets (PLT), 2.4 U; fresh frozen plasma (FFP), 7.5 U; and cryoprecipitate, 0 U. Explantation data on 134 patients showed a mean use as follows: RBC, 7.7 U; PLT, 2.4 U; FFP, 9.9 U; and cryoprecipitate, 3.7 U. Our protocol for the preparation of blood components for implantation and explantation of a cardiac device is, implantation: RBC, 4 U; PLT, 1 U; FFP, 6 U; explantation: RBC, 8 U; PLT, 2 U; FFP, 8 U. Our data do not support the need for preparation of cryoprecipitate before implantation or explantation.

Cryoprecipitate-removed plasma 'cryo-removed plasma' as a source of factor IX in the treatment of haemophilia B.

Fresh frozen plasma (FFP) is the main source of factor IX (FIX) in the treatment of bleeding episodes of haemophilia B in the Philippines. Cryoprecipitate-removed plasma otherwise known in the Philippines as cryosupernate, is a by-product of cryoprecipitate preparation. These blood products expire in storage or are just thrown- away because of less demand for clinical use. By theory, this product should have almost the same amount of FIX as in FFP, therefore can be used in the treatment of haemophilia B. There is no local data on the actual FIX content of the cryoprecipitate-removed plasma. Hence, the authors established these data to support the use of this product. Eighty-three bags of cryoprecipitate-removed plasma received from three different blood banks in Manila, Philippines were tested for FIX activity using an activated partial thromboplastin time (APTT)-based one-stage FIX assay. The FIX content in each bag of cryoprecipitate-removed plasma was calculated by multiplying its volume in mL with that of FIX activity per mL of plasma measured in vitro. The total mean FIX content per bag was 212.20 U (+/-88.98) exceeding the contents set by the American Association of Blood Banks (AABB, 70-90 U). The mean FIX activity per bag was 127.62% (+/-38.23) with the mean volume of 164.28 mL (+/-52.23). Statistically significant difference on volume (P = 0.000) was found across the three sources resulting to a significant variation of the actual FIX content (P = 0.000).

Desmopressin (DDAVP) and factor VIII: the tale as viewed from Milan (and Malmö)

Desmopressin (DDAVP) and factor VIII: the tale as viewed from Milan (and Malmö)

Proteolysis of von Willebrand Factor in Therapeutic Plasma Concentrates

Therapeutic plasma concentrates containing vo Willebrand factor (vWF) lack the largest, most hemostatically active multimers. To evaluate whether this abnormality results from proteolysis during manufacturing, we have analyzed the subunit structure of vWF in several commercial products and found a marked reduction in the relative content of intact 225-kD subunit, paralleled by an increase in the proteolytic fragments normally present in plasma, particularly that of 176 kD. There was no heightened vWF fragmentation in blood-bank cryoprecipitate prepared from platelet-poor, single-donor plasma; in contrast, there was a marked degree of fragmentation in cryoprecipitate prepared from pooled plasmapheresis plasma representing the starting fraction for the production of commercial concentrates. In cryoprecipitate prepared experimentally from plasma containing varying numbers of platelets, the degradation of vWF was proportional to the platelet count, but was greatly diminished by adding protease inhibitors to the plasma. On the basis of these findings, we postulate that the loss of the largest vWF multimers in commercial products results from the use of poorly centrifuged plasmapheresis plasma containing an excessive number of residual platelets and leukocytes. These cells, lysing when plasma is frozen and thawed for the preparation of cryoprecipitate, may liberate proteolytic enzymes that cleave the vWF subunit and contribute to the degradation of the largest multimers. Our results should help devise new approaches for the preparation of more effective concentrates for the treatment of von Willebrand disease.

Influence of heat treatment on FVIII:C recovery from freeze dried cryoprecipitate.

A standard lyophilised triple cryoprecipitate preparation, stabilised by the addition of Synthamin 17, was heat treated at 60 degrees C for 48 hours. The total protein content, factor VIII concentration, and factor VIII recovery were not affected by the heat treatment procedure. Heat treatment did not influence the reconstitution characteristics of the freeze dried preparation and there were no side effects during or after administration. The mean in vivo rise of factor VIII from infused heat treated triple cryoprecipitate was 2.5 (SD 0.9)%/unit/kg with a half life of 13.1 (3.1) hours. These results compare favourably with those obtained using non-heated triple cryoprecipitate. Cryoprecipitate can be heat treated without adversely influencing factor VIII recovery, and the ability to prepare a heat treated cryoprecipitate means that a small pool high yield factor VIII preparation can again be used in routine clinical practice.

Comparison of alpha-2-opsonic protein concentrations in cryoprecipitate prepared by methods of standard centrifugation and thaw-siphon techniques

Alpha-2-surface-binding glycoprotein (A2SBG) in cryoprecipitate prepared by standard centrifugation and thaw-siphon techniques was compared. The protein was quantitated by electroimmunoassay. The yield of A2SBG in the cryoprecipitate prepared by thaw-siphon (6.2 mg/mL) was significantly higher (P ≤ 0.01) than that prepared by standard centrifugation (3.9 mg/mL). Thaw-siphon cryoprecipitate preparation appears more cost effective in the treatment of patients with opsonic protein deficiency.