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CLC number: Q28

On-line Access: 2013-07-04

Received: 2012-08-22

Revision Accepted: 2013-05-28

Crosschecked: 2013-06-19

Cited: 2

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Journal of Zhejiang University SCIENCE B 2013 Vol.14 No.7 P.596-603


Complete disassociation of adult pancreas into viable single cells through cold trypsin-EDTA digestion

Author(s):  Dan Li, Shi-yun Peng, Zhen-wu Zhang, Rui-cheng Feng, Lu Li, Jie Liang, Sheng Tai, Chun-bo Teng

Affiliation(s):  Laboratory of Animal Development Biology, College of Life Science, Northeast Forestry University, Harbin 150040, China; more

Corresponding email(s):   taishengster@gmail.com, chunboteng@nefu.edu.cn

Key Words:  Mouse, Pancreas, Disassociation, Single cells, Viability, Cold digestion

Dan Li, Shi-yun Peng, Zhen-wu Zhang, Rui-cheng Feng, Lu Li, Jie Liang, Sheng Tai, Chun-bo Teng. Complete disassociation of adult pancreas into viable single cells through cold trypsin-EDTA digestion[J]. Journal of Zhejiang University Science B, 2013, 14(7): 596-603.

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journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

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%T Complete disassociation of adult pancreas into viable single cells through cold trypsin-EDTA digestion
%A Dan Li
%A Shi-yun Peng
%A Zhen-wu Zhang
%A Rui-cheng Feng
%A Lu Li
%A Jie Liang
%A Sheng Tai
%A Chun-bo Teng
%J Journal of Zhejiang University SCIENCE B
%V 14
%N 7
%P 596-603
%@ 1673-1581
%D 2013
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1200226

T1 - Complete disassociation of adult pancreas into viable single cells through cold trypsin-EDTA digestion
A1 - Dan Li
A1 - Shi-yun Peng
A1 - Zhen-wu Zhang
A1 - Rui-cheng Feng
A1 - Lu Li
A1 - Jie Liang
A1 - Sheng Tai
A1 - Chun-bo Teng
J0 - Journal of Zhejiang University Science B
VL - 14
IS - 7
SP - 596
EP - 603
%@ 1673-1581
Y1 - 2013
PB - Zhejiang University Press & Springer
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DOI - 10.1631/jzus.B1200226

The in vitro isolation and analysis of pancreatic stem/progenitor cells are necessary for understanding their properties and function; however, the preparation of high-quality single-cell suspensions from adult pancreas is prerequisite. In this study, we applied a cold trypsin-ethylenediaminetetraacetic acid (EDTA) digestion method to disassociate adult mouse pancreata into single cells. The yield of single cells and the viability of the harvested cells were much higher than those obtained via the two commonly used warm digestion methods. Flow cytometric analysis showed that the ratio of ductal or BCRP1-positive cells in cell suspensions prepared through cold digestion was consistent with that found in vivo. Cell culture tests showed that pancreatic epithelial cells prepared by cold digestion maintained proliferative capacity comparable to those derived from warm collagenase digestion. These results indicate that cold trypsin-EDTA digestion can effectively disassociate an adult mouse pancreas into viable single cells with minimal cell loss, and can be used for the isolation and analysis of pancreatic stem/progenitor cells.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


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