Cryopreservation is a desirable method for the long-term storage of human red blood cells (RBCs). Current protocols employ high concentrations of glycerol that must be removed from thawed RBCs prior to transfusion. Small-molecule ice recrystallization inhibitors (IRI) can protect RBCs from cryoinjury during the freezing and thawing process in the presence of reduced amounts of glycerol. Although reducing the concentration of glycerol during freezing reduces post-thaw deglycerolization times, thawed RBC units still require post-thaw processing. Herein, we report the cryopreservation of RBCs using the non-permeating cryoprotective agent (CPA) hydroxyethyl starch (HES) supplemented with small-molecule IRIs: (1) PMP-Glc (110 mM); and (2) pBrPh-Glc (30 mM). The results demonstrate that 30 mM pBrPh-Glc in 11.5 % (w/w) HES affords quantitative post-thaw recovery of intact RBCs that are superior to those obtained using glycerol with slow cooling rates, and show the utility of small-molecule IRIs in cryopreservation…
Category: <span>Cell & Tissue Banking</span>
Biological samples such as human tissue, cell lines, and serum specimens are a direct source of molecular data that can be used to identify targets for disease therapy, detection, and prevention. However, the validity and reliability of data derived from burgeoning analytical technology is highly dependent upon properly preserved, well-annotated biospecimens. As a result, biopreservation and biobanking have become an increasingly integral component in advancing today’s biotherapeutic development…
Storage and shipping variables may significantly impact cryopreserved product quality. In this particular article, the focus is on temperature control issues. The AATB (American Association of Tissue Banks) Standards for Tissue Banking state that cryopreserved cardiac allografts shall be maintained at temperatures of –100°C or colder. Heart valves are usually stored below –135°C in vapor phase nitrogen. There have been few published studies of higher (warmer) storage temperatures. Most heart valves are transported using dry shippers that maintain vapor phase nitrogen temperatures. These containment devices are expensive, and the costs for two-way shipping are significant due to their size and weight. Considerable savings could be had if dry ice shippers with temperature excursions warmer than the AATB standard (of up to –70°C) could be employed…
Three types of heart valves are employed for replacement in patients: mechanical, xenogeneic tissue, and allogeneic human valves derived from donors post-mortem. The intention of this article is two-fold: 1) to place issues associated with the preservation of allogeneic human heart valves in perspective relative to the AATB Standards for Tissue Banking and current practices in US cardiovascular tissue-processing facilities; and 2) present potential opportunities for improvements with appropriate validation. Most patients receive either xenogeneic tissue or mechanical valves; however, the use of cryopreserved human valve allografts became established during the 1970s and 1980s for certain patient subsets…
The enzyme-linked immunospot (ELISpot) assay is one of the most useful techniques for the immunological monitoring of vaccine trials and has increasing application as a measure of specific T-cell activation. Recently, we developed, optimized, and validated a customized ELISpot kit for the detection of interferon gamma (IFNγ) positive cells. The precision of the ELISpot was good and it varied over the range of the assay values, independent of the stimulus. Here we describe the development of a library of donors with characterized responses to the CEF peptide pool: cytomegalovirus (CMV), Epstein-Barr virus (EBV), and influenza (Flu); pool of 32 peptides which can be used as controls for IFNγ ELISpot and multiple immune monitoring assay validations for use in clinical trials…
The modern age of blood transfusion began after the Second World War, as detailed in Douglas Starr’s book, Blood: An Epic History of Medicine and Commerce. During the war, it became apparent that early and aggressive medical treatment utilizing whole blood or plasma could increase the chances of survival for military personnel wounded in combat. In the United States, a national program to encourage blood donation was created to provide the needed blood, which was then shipped as whole blood or plasma to war zones. After the war, physicians were eager to apply surgical advances developed on and off the battlefield to the care of the general population. Because these advances relied on blood transfusion, for the public to realize their benefit, adequate supplies of whole blood and blood components needed to be available to hospitals across the country. This was often not the case…
Within the United States, greater than 90% of the available transfusible blood products are collected, processed, and distributed by regional blood centers. The remaining blood products are collected by hospital-based blood banks and are usually provided only to patients in the collecting facility. The “region” in which a blood center offers services (i.e., collecting blood from volunteer donors and providing blood components to healthcare facilities), is usually an arbitrarily and independently defined group of contiguous counties surrounding a major metropolitan area. However, the borders of the region can be elastic and easily altered by gaining or losing access to donor groups or customers. It is not uncommon for a geographic area to be simultaneously “claimed” by neighboring competing blood centers. America’s Blood Centers (ABC) is a not-for-profit trade organization that provides services and advocacy for independent not-for-profit regional blood centers. ABC’s seventy-five members collect approximately 45% of the country’s blood products…