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Isolation, cultivation and identification of pancreatic stem/progenitor cells in rabbit

Published online by Cambridge University Press:  12 February 2007

Zhang Hui-Ru
Affiliation:
Shaanxi Branch Center of China Stem Cell Research Center, Northwest Sci-tech University of Agriculture and Forestry, Yangling, Shaanxi 712100, China College of Bioengineering, Henan University of Technology, Zhengzhou, Henan 450052, China
Feng Ruo-Peng
Affiliation:
Shaanxi Branch Center of China Stem Cell Research Center, Northwest Sci-tech University of Agriculture and Forestry, Yangling, Shaanxi 712100, China
Liu Xia
Affiliation:
Biology College of Yanan University, Yanan, Shaanxi 71600, China
Liu Yu-Xiao
Affiliation:
Shaanxi Branch Center of China Stem Cell Research Center, Northwest Sci-tech University of Agriculture and Forestry, Yangling, Shaanxi 712100, China Institution of Military Medical Science, Beijing 100850, China
Pang Quan-Hai
Affiliation:
Shaanxi Branch Center of China Stem Cell Research Center, Northwest Sci-tech University of Agriculture and Forestry, Yangling, Shaanxi 712100, China Shanxi Agricultural University, Taigu, Shanxi 030801, China
Dou Zhong-Ying*
Affiliation:
Shaanxi Branch Center of China Stem Cell Research Center, Northwest Sci-tech University of Agriculture and Forestry, Yangling, Shaanxi 712100, China
*
*Corresponding author. Email: douzhongying@china.com

Abstract

Pancreases obtained from 1–9-day-old rabbits were digested with 1 mg/ml ?? collagenase and cultivated with low-glucose Dulbecco's modified Eagle's medium (DMEM). The cells were subcultured almost until confluence. They were identified as pancreatic stem/progenitor cells using two methods: an immunohistochemical staining technique and the response of glucose-stimulated insulin secretion after β-cell induction. Results showed that the cells of rabbits acquired strong proliferation ability, and expressed the markers of pancreatic stem/progenitor cells: PDX-1, CK19 and nestin. After β-cell induction for 2 days, 20% of cells were dithizone (DTZ) positive, rapidly increasing to 80% after 6 days. Insulin secretion was stimulated by a high glucose concentration (26 mmol/l). These powerful tests demonstrated that isolated and cultivated cells in rabbits were pancreatic stem/progenitor cells. This result has not yet been reported elsewhere.

Type
Research Article
Copyright
Copyright © China Agricultural University and Cambridge University Press 2006

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