Rapid purification of human ductal cells from human pancreatic fractions with surface antibody CA19-9

被引：35

作者：

Gmyr, V ^{[1
]}

Belaich, S ^{[1
]}

Muharram, G ^{[1
]}

Lukowiak, B ^{[1
]}

Vandewalle, B ^{[1
]}

Pattou, F ^{[1
]}

Kerr-Conte, J ^{[1
]}

机构：

[1] Univ Hosp Lille, INSERM, F-59045 Lille, France

来源：

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS | 2004年 / 320卷 / 01期

关键词：

human pancreatic ductal cell; cell therapy of diabetes; cell selection; neogenesis; surface marker; alternative source of insulin-secreting cells;

D O I：

10.1016/j.bbrc.2004.05.125

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：27 / 33

页数：7

共 32 条

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