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PLoS One. 2019 May 10;14(5):e0216136. doi: 10.1371/journal.pone.0216136. eCollection 2019.

Strategy towards tailored donor tissue-specific pancreatic islet isolation.

Author information

1
Department of Surgery, Tohoku University Graduate School of Medicine, Sendai, Miyagi, Japan.
2
Graduate School of Biomedical Engineering, Tohoku University, Sendai, Miyagi, Japan.
3
Division of Transplantation and Regenerative Medicine, Tohoku University Graduate School of Medicine, Sendai, Miyagi, Japan.
4
Department of Applied Biological Chemistry, Graduate School of Agricultural Science, Tokyo University of Agriculture and Technology, Fuchu, Tokyo, Japan.
5
New Industry Creation Hatchery Center, Tohoku University, Sendai, Miyagi, Japan.
6
Department of Biochemistry, Tohoku University Graduate School of Medicine, Sendai, Miyagi, Japan.

Abstract

BACKGROUND:

Optimizing 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.

METHODS:

Islet 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.

RESULTS:

No 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.

CONCLUSIONS:

The 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.

Conflict of interest statement

The authors have declared that no competing interests exist.

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