Rapid purification of human ductal cells from human pancreatic fractions with surface antibody CA19-9.
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.
Gmyr
Valéry
V
Cell Therapy of Diabetes, INSERM ERIT-M 0106, University Hospital of Lille, 1 place de Verdun, Lille 59045, France.
Belaich
Sandrine
S
Muharram
Ghaffar
G
Lukowiak
Bruno
B
Vandewalle
Brigitte
B
Pattou
François
F
Kerr-Conte
Julie
J
D018395:CA-19-9 Antigen / Q000276:immunology*; D002478:Cells, Cultured; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods*; D007515:Islets of Langerhans / Q000166:cytology* / Q000502:physiology*; D010179:Pancreas / Q000166:cytology* / Q000502:physiology*; D010183:Pancreatic Ducts / Q000166:cytology / Q000502:physiology
D018395:CA-19-9 Antigen