JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES) ›› 2012, Vol. 50 ›› Issue (3): 8-.

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Improved method for rat pancreatic islets isolation and purification

XIAO Fang1,2,3, LI Wen-juan1,2,3, HOU Xin-guo1,2, SONG Jun1,2, XIAO Juan1,2, WANG Mei-jian1,2,
LIU Ji-dong1,2, DENG Nan1,2, ZHOU Xiao-ying1,2, CHEN Li1,2   

  1. 1. Institute of Endocrinology and Metabolism; 2. Department of Endocrinology, Qilu Hospital;
    3. Centre for Cell Therapy Research, Shandong University, Jinan 250012, China
  • Received:2011-10-13 Online:2012-03-10 Published:2012-03-10

Abstract:

Objective   To optimize conditions for rat pancreatic islets isolation and purification, and provide experimental data for research on the pathogenesis of islet injury in diabetes and application of islet regeneration. Methods   Influences of different pancreatic perfusion volumes and digestion time on rat pancreatic islets isolation through intraductal collagenase V perfusion in situ were observed.  Effects on islets purification were compared between classical discontinuous Ficoll density gradient centrifugation and Histopaque 1077 density gradient centrifugation. Afterwards, the islets were examined and their morphology, quantity, purity and function were evaluated in vitro.  Results   Histopaque 1077 density gradient centrifugation could yield more islets of high quality (P<0.01). Insulin release by the islets through the two different methods under high glucose condition has no differentiation(P>0.05). Islets perfusion volumes and digestion time played significant roles in rat islets extraction. Conclusion   Histopaque 1077 density gradient centrifugation, with simple operation and high yield, has a prospect to become the most effective method for rat islets purification and it will be more useful in clinical and basic research in the future.

Key words: Rat islets; Isolation; Purification; Collagenase V

CLC Number: 

  • R587.1
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