PLoS ONE (Jan 2019)

Strategy towards tailored donor tissue-specific pancreatic islet isolation.

  • Yuki Miyazaki,
  • Kazutaka Murayama,
  • Ibrahim Fathi,
  • Takehiro Imura,
  • Youhei Yamagata,
  • Kimiko Watanabe,
  • Hiroshi Maeda,
  • Akiko Inagaki,
  • Yasuhiro Igarashi,
  • Shigehito Miyagi,
  • Hiroki Shima,
  • Kazuhiko Igarashi,
  • Takashi Kamei,
  • Michiaki Unno,
  • Masafumi Goto

DOI
https://doi.org/10.1371/journal.pone.0216136
Journal volume & issue
Vol. 14, no. 5
p. e0216136

Abstract

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BackgroundOptimizing the collagenase G (ColG):collagenase H (ColH) ratio is a key strategy for achieving tailored donor-tissue specific islet isolation. Collagen V (Col V) and collagen III (Col III) are crucial target matrices of ColG and ColH, respectively. We herein investigated the relevance between the expression of target matrices in pancreatic tissues and influence of ColG:ColH ratio on islet isolation outcome.MethodsIslet isolation was performed in Lewis and SD rats using different ColG:ColH ratios (5:1, 1:1 and 1:5; n = 7/group). The composition of Col III and Col V was examined using immunohistochemical staining, real-time polymerase chain reaction (PCR), Western blotting and mass spectrometry. Chain types in collagen I (Col I) were also assessed using mass spectrometry.ResultsNo beneficial effects were observed by increasing the ColG amount, irrespective of the rat strain. In contrast, the islet yield in Lewis rats was considerably increased by high amounts of ColH but decreased in SD rats, suggesting that Lewis pancreas contains more Col III than SD pancreas. Neither immunohistochemical nor real-time PCR showed correlation with isolation outcome. However, Western blotting revealed that Lewis contained considerably higher amount of Col III than SD (p = 0.10). Likewise, Col-I(α1)/Col-III(α1) and Col-I(α2)/Col-III(α1) were significantly lower in Lewis than in SD rats (p = 0.007, respectively). Furthermore, the isolation outcome was considerably correlated with the composition of homotrimeric Col I.ConclusionsThe Col III expression and the composition of homotrimeric Col I in pancreatic tissues determined using mass analyses appeared useful for optimizing the ColG:ColH ratio in islet isolation.