STUDY OF SURVIVAL AND FUNCTIONALITY IN CRYOPRESERVED PLATELETS

SURVIVAL AND FUNCTIONALITY EVALUATION IN VITRO IN CRYOPRESERVED PLATELETS

Platelets correspond to one of the most widely used blood components in transfusion medicine, given their importance in the prevention of bleeding disorders in patients with multifactorial thrombocytopenia and non-immunological etiology, as well as in the management of traumatic or blood-related bleeding. surgical treatments. Given its wide margin of use and the constant demand for it for use within the different management schemes and procedures associated with the medical act, it is the objective of both hemotherapy services and blood banks and blood products to have availability component constant to meet such requirements. The main problem of this resource is its short useful life (5 to 7 days) which limits its availability, especially in the Latin American context where the production of blood components as well as the resources allocated for this purpose by state entities to blood banks are generally limited. Cryopreservation at -80 ° C using Dimethylsulfoxide as a preservative solution is a technique used since the 1950s to prolong platelet survival, the improvement of which has been remarkable in recent years, thus constituting an alternative for the resolution of this problem.

Study Overview

• Status: Recruiting
• Conditions: Platelet Cryopreservation; Platelet Survival
• Intervention / Treatment: Other: platelet cryopreservation

Detailed Description

HYPOTHESIS: Through cryopreservation it is possible to prolong the survival time of platelets up to 4 times, compared to those preserved under the conventional method, maintaining a viability equal to or greater than 50%.

There is a direct relationship between survival time, viability and functionality of cryopreserved platelets depending on the model of cryoprotective solution used.

PRIMARY OUTCOME: survival of cryopreserved platelets greater than 28 days SECUNDARY OUTCOME: viability of cryopreserved platelets greater than 50%

INCLUSION CRITERIA: Concentrates from donors over 18 years of age who meet the requirements established by law 22990 (Of Argentine Republic) for donating blood and blood components will be included in this study, from whom a number of platelets greater than or equal to 3 x10 E11 can be collected.

EXCLUSION CRITERIA: Platelet concentrates that present any of the following characteristics will be excluded from the investigation:

Concentrates that have not met all of the standards required for use in humans or that at the time of the study more than 24 hours have passed since they were obtained.

A blind experimental study will be carried out (those who evaluate viability and functionality of platelets will not know the cryopreservation times or the concentration of the cryopreservative)

Aliquots of platelets with group and factor A +, A-, B +, B-, AB +, AB-, O + and O- obtained from those people who come to the hemotherapy center as voluntary donors and who in turn express their decision to participate in this protocol by signing an informed consent designed for this purpose. Obtaining platelets will be performed by apheresis using the Trima ACCEL cell separator, version 7.0, TERUMO BCT brand.

A minimum total of 3.3 x1011 platelets will be obtained in a volume between 200 - 300 ml, for each donor.

Group A: It will consist of 70 aliquots of 3 ml each with 1.9 x109 platelets to which a solution composed of: 1 ml obtained from the following mixture will be added (0.8 ml of 100% Dimethylsulfoxide (DMSO) and 4 ml of 0 sodium chloride , 9%). They will then be frozen at -80 ° C.

B Group: It will be made up of 70 aliquots of 3 ml each with 1.9 x109 platelets to which will be added a solution composed of: 0.4 ml of 20% UNC brand albumin and 0.6 ml obtained from the following mixture (1.4 ml of Dimethylsulfoxide (DMSO) 100% and 4 ml of Dextrose 5%). They will then be frozen at -80 ° C.

Group C: It will be made up of 70 aliquots of 3 ml each with 1.9 x109 platelets to which will be added a solution composed of: 0.4 ml of 20% UNC brand albumin and 0.6 ml obtained from the following mixture (1.4 ml of Dimethylsulfoxide (DMSO) 100% and 4 ml of 0.9% sodium chloride). They will then be frozen at -80 ° C.

A group of 71 aliquots of 3 ml each with 1.9 x109 irradiated platelets kept at 22 ° C under constant stirring for 5 days (twenty-four aliquots for each period of time evaluated) will be used as a reference for the comparison of results (group N).

Viability:

It will be evaluated through the platelet count carried out by the Beckman-Coulter system, the measurement of the pH of the cryopreserved platelets after being thawed (expected values between 6.4 and 7.4) and the immunophenotyping of the platelets by flow cytometry (CMF) using fluorescent anti CD61-PE antibodies.

Functionality:

It will be evaluated from the immunophenotyping of platelets by flow cytometry (CMF) using fluorescent anti-CD41 and anti-CD62-FITC antibodies.

Before carrying out the CMF analysis, 4 microliters of a 50 mmol ADP solution will be added to each sample in order to induce platelet activation.

Both the viability analysis and the functionality of the cryopreserved product will be carried out by 3 biochemists, collaborating members of the research in the biochemical analysis laboratory of the hospital.

• Study Type: Interventional
• Enrollment (Anticipated): 281
• Phase: Not Applicable

Contacts and Locations

• Study Contact: Name: JHON AVILA; Phone Number: +5491141879723; Email: jaavilar@gmail.com
• Study Contact Backup: Name: JOSE ACOSTA; Email: jacosta@fundaleu.org.ar

Study Locations

• Argentina
  • Buenos Aires, Argentina, 1082
    • Recruiting
    • Hospital Britanico Buenos Aires
    • Contact: JHON AVILA, MD; Phone Number: +5491141879723; Email: jaavilar@gmail.com
  • Buenos Aires, Argentina, 1406
    • Completed
    • Hospital Británico de Buenos Aires

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years and older (ADULT, OLDER_ADULT)
• Accepts Healthy Volunteers: Yes
• Genders Eligible for Study: All

Description

Inclusion Criteria:

• Concentrates from donors over 18 years of age who meet the requirements established by law 22990 for donating blood and blood components will be included in this study, from whom a number of platelets greater than or equal to 3 x10 E11 can be collected

Exclusion Criteria:

• Platelet concentrates that present any of the following characteristics will be excluded from the investigation:

Concentrates that have not met all of the standards required for use in humans or that at the time of the study more than 24 hours have passed since they were obtained.

Study Plan

How is the study designed?

Design Details

• Primary Purpose: BASIC_SCIENCE
• Allocation: NA
• Interventional Model: SINGLE_GROUP
• Masking: NONE

Number of Arms

1

Arms and Interventions

Participant Group / Arm

Intervention / Treatment

EXPERIMENTAL: SURVIVAL AND FUNCTIONALITY INVITRO IN CRYOPRESERVED PLATELETS; Platelet concentrates will be administered 3 alternatives of cryopreservative solution: 5% dimethylsulfoxide (solution 1), 5% dimethylsulfoxide plus 160 mg of 5% dextrose (solution 2) and 5% dimethylsulfoxide plus 2% albumine (solution 3 ). They will then be frozen at -80 ° C and their survival and functionality will be subsequently evaluated in vitro.

Other: platelet cryopreservation; The cryopreservation of platelets will be carried out using as alternatives of cryopreservative solution: 5% dimethylsulfoxide alone (solution 1) or combined with 5% dextrose 160 mg (solution 2) or with 2% albumin (solution 3) and then be frozen at -80 ° C

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Time Frame
survival of cryopreserved platelets greater than 28 days	Between 1 and 3 months

Secondary Outcome Measures

Outcome Measure	Time Frame
viability of cryopreserved platelets greater than 50%	Between 1 and 3 months

Collaborators and Investigators

• Sponsor: HOSPITAL BRITANICO DE BUENOS AIRES
• Investigators: Study Chair: GLENDA ERNST, Hospital Británico de Buenos Aires; Principal Investigator: JHON AVILA, Hospital Británico de Buenos Aires

Publications and helpful links

General Publications

• Johnson L, Cameron M, Waters L, Padula MP, Marks DC. The impact of refrigerated storage of UVC pathogen inactivated platelet concentrates on in vitro platelet quality parameters. Vox Sang. 2019 Jan;114(1):47-56. doi: 10.1111/vox.12730. Epub 2018 Nov 30.
• 7. Bohonek, Milos;Cryopreservation of Platelets: Advances and Current Practice; Cryopreservation Biotechnology in Biomedical Sciences ;2018;Chapter4;P47-66
• Waters L, Cameron M, Padula MP, Marks DC, Johnson L. Refrigeration, cryopreservation and pathogen inactivation: an updated perspective on platelet storage conditions. Vox Sang. 2018 May;113(4):317-328. doi: 10.1111/vox.12640. Epub 2018 Feb 14.
• Eker I, Yilmaz S, Cetinkaya RA, Pekel A, Unlu A, Gursel O, Yilmaz S, Avcu F, Musabak U, Pekoglu A, Ertas Z, Acikel C, Zeybek N, Kurekci AE, Avci IY. Generation of Platelet Microparticles after Cryopreservation of Apheresis Platelet Concentrates Contributes to Hemostatic Activity. Turk J Haematol. 2017 Mar 1;34(1):64-71. doi: 10.4274/tjh.2016.0049. Epub 2016 Apr 18.

Study record dates

Study Major Dates

• Study Start (ACTUAL): February 3, 2021
• Primary Completion (ANTICIPATED): August 28, 2021
• Study Completion (ANTICIPATED): February 2, 2022

Study Registration Dates

• First Submitted: February 7, 2021
• First Submitted That Met QC Criteria: February 11, 2021
• First Posted (ACTUAL): February 15, 2021

Study Record Updates

• Last Update Posted (ACTUAL): August 3, 2021
• Last Update Submitted That Met QC Criteria: July 28, 2021
• Last Verified: July 1, 2021

More Information

Terms related to this study

• Keywords: IN VITRO; SURVIVAL; FUNCTIONALITY; CRYOPRESERVED PLATELETS
• Other Study ID Numbers: 1021

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No