Abstract 448: Isolation and Characterization of Lesion Specific Cells Harvested in vivo During Percutaneous Coronary Intervention

Abstract

Background: Endothelial cells (EC) dysfunction is linked to the development of CAD. Current studies rely on ECs harvested from the umbilical vein or bovine aorta because of the ease of isolation. This fails to account for phenotypic variations and may not adequately represent pathologic changes occurring at the site of specific coronary artery lesion. The need to examine EC phenotypes and cellular processes in the context of existing disease is necessary. Hypothesis: Coronary artery lesion specific cells can be harvested and isolated from baloons used during PCI. Methods: Coronary balloons used during PCI were immediately placed in endothelial cell media and subjugated to a series of washes, including phosphate buffered saline (PBS) and trypsin. The washes and balloons were centrifuged at 1,700 rpm for 10 minutes and the resulting pellets re-suspended in ammonium-chloride-potassium (ACK)/PBS solution to lyse erythrocytes. These suspensions were centrifuged at 2,500 rpm for 10 minutes to pellet the cells. An aliquot of this final cell harvest was analyzed by fluorescence- activated cell sorting (FACS) with antibodies for endothelial antigens. Results: 30 samples were analyzed and subpopulations of cells were identified. Between 400-4000 non-hematologic cells per sample were isolated. Cell populations were considered to be endothelial when they were CD31, CD146 double positive and CD45 negative, and populations that were CD222 positive (CD56 CD45 double negative) and CD56 positive (CD222 and CD45 negative) were considered to be fibroblastic and myogenic respectively. Endothelial cells (Figure 1A), myogenic cells (Figure 1B) and fibroblastic cells (Figure 1C) could be visualized. Conclusions: Lesion specific cells were successfully harvested and isolated from coronary balloons used during PCI. Proteomic and genetic analyses of these cells may provide invaluable information about pathologic cellular processes during ACS.