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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 ,
Feng Ruo-Peng ,
Liu Xia ,
Liu Yu-Xiao ,
Pang Quan-Hai  and
Dou Zhong-Ying
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
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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.

Keywords

identificationinsulin-secretionpancreatic stem cellsrabbit
Type
Research Article
Information
Chinese Journal of Agricultural Biotechnology , Volume 3 , Issue 2 , August 2006 , pp. 89 - 94
Copyright
Copyright © China Agricultural University and Cambridge University Press 2006

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References

Abraham, JE, Leech, AC, Lin, CJ, Zulewski, H and Habener, FJ (2002) Insulinotropic hormone glucagon-like peptide-1 differentiation of human pancreatic islet-derived progenitor cells into insulin-producing cells. Endocrinology 143(8): 3152–3161.CrossRefGoogle ScholarPubMed
Basta, G, Racanicchi, L, Mancuso, F, et al. (2004) Neonatal pig pancreatic duct–derived insulin-producing cells: preliminary in vitro studies. Transplantation Proceedings 36: 609–611.CrossRefGoogle ScholarPubMed
Blyszczuk, P and Wobus, AM (2004) Stem cells and pancreatic differentiation in vitro. Journal of Biotechnology 113: 3–13.CrossRefGoogle ScholarPubMed
Cebrian, A, García-Ocaña, A, Takane, KK, Sipula, D, Stewart, AF and Vasavada, RC (2002) Overexpression of parathyroid hormone-related protein inhibits pancreatic β-cell death in vivo and in vitro. Diabetes 51: 3003–3013.CrossRefGoogle ScholarPubMed
Colman, A (2004) Making new beta cells from stem cells. Seminars in Cell and Developmental Biology 15: 337–345.CrossRefGoogle ScholarPubMed
Garcia-Ocan, A, Takane, KK, Syed, MA, Philbrick, WM, Vasavada, RC and Stewart, AF (2000) Hepatocyte growth factor overexpression in the islet of transgenic mice increases beta cell proliferation, enhances islet mass, and induces mild hypoglycemia. Journal of Biological Chemistry 275(2): 1226–1232.CrossRefGoogle Scholar
Gmyr, V, Kerr-Conte, J, Belaich, S, et al. (2000) Adult human cytokeratin 19–positive cells reexpress insulin promoter factor 1 in vitro. Diabetes 49: 1671–1680.CrossRefGoogle ScholarPubMed
Hosotani, R, Ida, J, Kogire, M, Fujimoto, K, Doi, R and Imamura, M (2004) Expression of pancreatic duodenal hoemobox-1 in pancreatic islet neogenesis after surgical wrapping in rats. Surgery 135: 297–306.CrossRefGoogle ScholarPubMed
Kojima, H, Nakamura, T, Fujita, Y, et al. (2002) Combined expression of pancreatic duodenal homeobox 1 and islet factor 1 induces immature enterocytes to produce insulin. Diabetes 51: 1398–1408.CrossRefGoogle ScholarPubMed
Ohgawara, H, Shikano, T, Fukunaga, K, Yamagishi, M and Miyazaki, H (1998) Establishment of monolayer culture of pig pancreatic endocrine cells by use of nicotinamide. Diabetes Research and Clinical Practice 42: 1–8.CrossRefGoogle ScholarPubMed
Sato, A, Wang, P-C and Ohgawara, H (2002) Effect of stimulators such as GLP-1, PACAP, and nicotinamide on glucose-stimulated insulin secretion from porcine pancreatic endocrine cells in long-term culture. Pancreas 25(1): 86–93.CrossRefGoogle ScholarPubMed
Shiroi, A, Yoshikawa, M, Yokota, H, et al. (2002) Identification of insulin-producing cells derived from embryonic stem cells by zinc-chelating dithizone. Stem Cells 20: 284–292.CrossRefGoogle ScholarPubMed
Tour, DD, de la Halvorsen, T, Demeterco, C, et al. (2001) β-Cell differentiation from a human pancreatic cell line in vitro and in vivo. Molecular Endocrinology 15(3): 476–483.Google ScholarPubMed
Wang, R, Yashpal, N, Bacchus, F and Li, J (2004) Hepatocyte growth factor regulates proliferation and differentiation of epithelial monolayers derived from islets of postnatal rat pancreas. Journal of Endocrinology 183: 163–171.CrossRefGoogle ScholarPubMed