Shipping blood to a central laboratory in multicenter clinical trials: effect of ambient temperature on specimen temperature, and effects of temperature on mononuclear cell yield, viability and immunologic function

Abstract

BackgroundClinical trials of immunologic therapies provide opportunities to study the cellular and molecular effects of those therapies and may permit identification of biomarkers of response. When the trials are performed at multiple centers, transport and storage of clinical specimens become important variables that may affect lymphocyte viability and function in blood and tissue specimens. The effect of temperature during storage and shipment of peripheral blood on subsequent processing, recovery, and function of lymphocytes is understudied and represents the focus of this study.MethodsPeripheral blood samples (n = 285) from patients enrolled in 2 clinical trials of a melanoma vaccine were shipped from clinical centers 250 or 1100 miles to a central laboratory at the sponsoring institution. The yield of peripheral blood mononuclear cells (PBMC) collected before and after cryostorage was correlated with temperatures encountered during shipment. Also, to simulate shipping of whole blood, heparinized blood from healthy donors was collected and stored at 15°C, 22°C, 30°C, or 40°C, for varied intervals before isolation of PBMC. Specimen integrity was assessed by measures of yield, recovery, viability, and function of isolated lymphocytes. Several packaging systems were also evaluated during simulated shipping for the ability to maintain the internal temperature in adverse temperatures over time.ResultsBlood specimen containers experienced temperatures during shipment ranging from -1 to 35°C. Exposure to temperatures above room temperature (22°C) resulted in greater yields of PBMC. Reduced cell recovery following cryo-preservation as well as decreased viability and immune function were observed in specimens exposed to 15°C or 40°C for greater than 8 hours when compared to storage at 22°C. There was a trend toward improved preservation of blood specimen integrity stored at 30°C prior to processing for all time points tested. Internal temperatures of blood shipping containers were maintained longer in an acceptable range when warm packs were included.ConclusionsBlood packages shipped overnight by commercial carrier may encounter extreme seasonal temperatures. Therefore, considerations in the design of shipping containers should include protecting against extreme ambient temperature deviations and maintaining specimen temperature above 22°C or preferably near 30°C.

Optimal cellular preservation for high dimensional flow cytometric analysis of multicentre trials

“…Studies conducted by Ahmann et al (2008) showed that tissue sample shipment on dry ice within USA did not have a drastic impact on plasma cell isolation, viability and RNA integrity. Olson et al (2011) had investigated the effects of overnight ambient temperature shipment at different seasons and samples subjected to 40°C for more than 8 h showed reduced viability. In Olson et al study, PBMCs were isolated for analysis and 4 color flow staining panel was employed.…”

Cardiovascular toxicity and titin cross-reactivity of affinity-enhanced T cells in myeloma and melanoma

“…8 Samples (peripheral blood, bone marrow) were collected in lavender-top (K 2 EDTA,) or red-top (no additive) BD vacutainer tubes (Becton Dickinson). Lavender-top tubes were delivered to the laboratory within 2 hours of draw or shipped overnight at room temperature in insulated containers essentially as described 9 prior to processing. Samples were processed within 30 minutes of receipt according to established laboratory standard operating procedures.…”

Preanalytical Variables Affecting the Integrity of Human Biospecimens in Biobanking

“…DBS should be transported to the laboratory at ambient temperature within 24 h of collection (19,32 ). Blood, urine, saliva, DNA, or RNA shipped by commercial carrier may encounter extreme seasonal temperatures, and this factor should thus be accounted for in long-term large-scale multinational studies (51,98,99 ). Samples that must be shipped to a processing laboratory cannot be used for the most unstable biomarkers, or processing should begin in the field to ensure stability during transportation (7 ).…”