Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (10): 1819-1822.doi: 10.3969/j.issn.1673-8225.2011.10.023

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A method of pancreatic stem cells culture and amplification in vitro 

Kou Ya-li1, Hu Li-xia2, Li Qin1, Zhao Wen-jing1, Chen Wei-ping1   

  1. 1Department of Histology and Embryology, Guangxi Medical University, Nanning   530021,
    Guangxi Zhuang Autonomous Region, China; 2Department of Obstetrics, Xinxiang Central Hospital, Xinxiang  453000, Henan Province, China
  • Received:2010-10-14 Revised:2010-12-04 Online:2011-03-05 Published:2011-03-05
  • Contact: Chen Wei-ping, Professor, Master’s supervisor, Department of Histology and Embryology, Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China chwp8852@126.com
  • About author:Kou Ya-li★, Studying for master’s degree, Department of Histology and Embryology, Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China kouyali-2011@163.com
  • Supported by:

    Science and Technology Bureau of Guangxi Zhuang Autonomous Region, No.0229009*, 0592007-1G*, 0832157*

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Abstract:

BACKGROUND: Pancreatic stem cells are differentiated easily when cultured in vitro, so it is difficult to obtain a sufficient number of pancreatic stem cells.
OBJECTIVE: To explore a simple method which pancreatic stem cells were amplified and cultured in vitro.
METHODS: The pancreases of newborn Kunming mice were isolated. After culturing in vitro for 3-4 days, the colony growing pancreatic stem cells were emerged in fibroblasts. The number of fibroblasts was reduced by 0.02% EDTA; the fibroblasts were cultured continuously after their proportions were reduced. When the cells of confluent culture reached 80% of bottom, repeat 3 or 4 times, the number of pancreatic stem cells was gradually increased. The specific marker (Nestin) of pancreatic stem cells was detected. Differentiation of pancreatic stem cells was induced by nicotinamide. Islet-like cell clusters were detected by dithizone (DTZ) staining.
RESULTS AND CONCLUSION: After the number and proportion of fibroblasts were reduced, the pancreatic stem cells presented active proliferative ability. The number of the pancreatic stem cells was significantly increased. The amplificated pancreatic stem cells maintained their properties, including round appearance with single nucleus, strong diffraction, and expression (Nestin). After induced by nicotinamide, the amplificated pancreatic stem cells were differentiated to islet-like cell clusters with positive DTZ staining. The results demonstrated that the proportion and number of fibroblasts were reduced which were beneficial for proliferation of pancreatic stem cells. A large number of pancreatic stem cells were obtained. The amplificated pancreatic stem cells with undifferentition condition still had the potential of differentiation to pancreatic islet under suitable induction condition.

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