4.1
#79
CELL PREPARATION THAT CAN BE USED FOR SEROLOGICAL HLA TYPING, CROSSMATCHING, AND FLOW CROSSMATCHING.
Dale M. Dunn, MD , Mark N. Thurmon, BS and Cathy L. Sypert, BS . Lubbock TX, Texas Tech University Health Sciences Center, 79430, Department of Pathology .

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Most HLA laboratories prepare one cell prepartion for serological HLA typing and T- and B-Cell crossmathing and then prepare another cell preparation for Flow T- and B-Cell crossmatch. Development of the RosetteSep product by StemCell Technologies and distributed by Pel-Freez clinical systems provides a cell separation technique that can be used for both serological Class I and II HLA typing, crossmatching and T- and B-Cell Flow Crossmatching.
Many Histocompatibility laboratories use magnetic beads to harvest T- and B-Lymphocyte populations for CDC crossmatching, and HLA typing. And then they use a density gradient centrifugation method (Ficoll-Hypaque) to harvest lymphocytes and treat these cells with three-color reagents for T- and B-Cell Flow Crossmatching. The RosetteSep procedure creates an enriched lymphocyte cell preparation by rosetting out the unwanted cells, leaving populations of "untouched" T- or B-Lymphocytes. These cell preparations are approximately 98% pure for T-Cells and approximately 90% pure for B-Cells. They were tested in all the HLA applications listed above and found to give excellent results. Serological typing and CDC crossmatching plates were easy to read and interpret. The lack of magnetic beads created a less cluttered background and the cells show less nonspecific cell death than those separated with conventional magnetic bead technology. By analyzing the T- and B-Cells separately on the Flow Cytometer it significantly reduces the cost of performing the test and the labor required to set-up the instrument. RosetteSep methodology was easy to perform, cost effective, and utilized technologist time efficiently.