Generating human insulin-secreting cells for cell therapy of diabetes represents a highly competitive world challenge. Human ductal cells can give rise to islets in vivo and in vitro. The goal of this study was to devise a rapid sorting method to highly purify human ductal cells from pancreatic tissue using a pan-ductal membrane antibody carbohydrate antigen 19-9 (CA19-9). Human pancreatic sections confirmed antibody specificity. The human exocrine fraction (30% ductal cells) was sorted with magnetic bead technology or by FACS. Immunocytochemistry post-sorting determined ductal cell content. The manual magnetic bead technique resulted in 74% 2 (n = 4) CA19 positive cells. Whereas the automated AutoMACS technique (n = 5) yielded 92.6% +/- 0.5 CA19-9 positive cells with only a minor beta cell contamination (0.2% +/-0.03); cell yield post-sorting was 12.9% +/- 2.5 (1.69 +/- 0.41 x 10(6) cells) with 51.7% + 6.5 (n = 5) viability post-sorting. The FACS (n = 6) resulted in 97.1% +/- 0.82 CA19-9 positive cells, a cell yield of 25.5% +/- 5.6 (5.03 +/- 1.0 x 10(6)), with 72.1% +/- 6.1 viability post-sorting. (C) 2004 Elsevier Inc. All rights reserved.