Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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A Method for the Separation of Mouse Pancreatic Islets Using Discontinuous Percoll Gradient Centrifugation

  • Cho, Yu-Ree (Graduate School of Biotechnology, Korea University) ;
  • Kim, Sang-Duk (Graduate School of Biotechnology, Korea University) ;
  • Chang, Hyo-Ihl (Graduate School of Biotechnology, Korea University) ;
  • Sung, Ha-Chin (Graduate School of Biotechnology, Korea University) ;
  • Lee, Cherl-Ho (Graduate School of Biotechnology, Korea University) ;
  • Kim, Chan-Wha (Graduate School of Biotechnology, Korea University)
  • Published : 1999.08.01
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Abstract

A discontinuous Percoll gradient was used to separate islets from the collagenase-treated mouse pancreas easily and rapidly. Since the osmolality of Percoll is very low, adjustment of its osmolality to 340 mOs/kg $H_2$O was essential for securing the optimal separation. A discontinuous gradient layering with Percoll solution of 1.09 g, 1.07g, and1.05g/m, respectively, when centrifuged at 800$\times$g for 10 min, resulted in an optimal condition for separation and yielded a banding pattern with an even distribution of islet cells. No significant difference was observed in the morphological features between the Percoll-isolated and the manually-isolated islets. In conclusion, the discontinuous Percoll gradient can be effectively used to isolate the pancreatic islets from mice with four-fold higher efficiency compared to the handpicking method.

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References

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