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. 2014 Jul:409:99-106.
doi: 10.1016/j.jim.2014.05.019. Epub 2014 Jun 10.

Leukopak PBMC sample processing for preparing quality control material to support proficiency testing programs

Ambrosia Garcia  1 , Sarah Keinonen  1 , Ana M Sanchez  1 , Guido Ferrari  2 , Thomas N Denny  3 , M Anthony Moody  4
Affiliations

Leukopak PBMC sample processing for preparing quality control material to support proficiency testing programs

Ambrosia Garcia et al. J Immunol Methods. 2014 Jul.

Abstract

External proficiency testing programs designed to evaluate the performance of end-point laboratories involved in vaccine and therapeutic clinical trials form an important part of clinical trial quality assurance. Good clinical laboratory practice (GCLP) guidelines recommend both assay validation and proficiency testing for assays being used in clinical trials, and such testing is facilitated by the availability of large numbers of well-characterized test samples. These samples can be distributed to laboratories participating in these programs and allow monitoring of laboratory performance over time and among participating sites when results are obtained with samples derived from a large master set. The leukapheresis procedure provides an ideal way to collect samples from participants that can meet the required number of cells to support these activities. The collection and processing of leukapheresis samples require tight coordination between the clinical and laboratory teams to collect, process, and cryopreserve large number of samples within the established ideal time of ≤8 hours. Here, we describe our experience with a leukapheresis cryopreseration program that has been able to preserve the functionality of cellular subsets and that provides the sample numbers necessary to run an external proficiency testing program.

Keywords: Leukapheresis; Leukocytes; PBMC; Proficiency testing.

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Figures

Figure 1
Figure 1
Equipment used for the preparation, storage, and shipment of PBMC. A. Leukapheresis machine (Terumo BCT COBE Spectra Apheresis System). B. Hematology counter (Beckman Coulter Ac*T 5 Diff CP). C. Cell / viability counter (Beckman Coulter Vi-Cell XR). D. Controlled rate freezer (Gordinier Model #9010). E. Cryogenic cart (MVE CHART CryoCart™). F. Liquid nitrogen vapor phase storage tanks. G. Dry shipper container (MVE CHART); Dewar is inserted into this container. H. External shipping cover for dry shipper.
Figure 2
Figure 2
Leukopak processing. The steps required for correct and timely processing of a leukapheresis product are shown in chronological order.
Figure 3
Figure 3
Cryopreservation and storage of PBMC samples. A description of the requirements for long-term storage of the samples from time of collection is provided.
Figure 4
Figure 4
Shipping of cryopreserved samples. The steps required for proper distribution of the sample within country and internationally are provided. These steps have insured consistent quality (viability, recovery, and functionality) of the cryopreserved samples received by an end-point laboratory.
Figure 5
Figure 5
Sample selection screenshot. Sample selection is performed using a web-based interface that queries the LDMS record system. In the system, multiple samples can be selected and assembled into a shipping request. Prior to shipment, MTA and regulatory compliance is confirmed.
See this image and copyright information in PMC

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